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Immunoglobulins

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Marnila P and Korhonen H (2011) Milk Proteins | Immunoglobulins. In: Fuquay JW,
Fox PF and McSweeney PLH (eds.), Encyclopedia of Dairy Sciences, Second
Edition, vol. 3, pp. 807–815. San Diego: Academic Press.

ª 2011 Elsevier Ltd. All rights reserved.

 Author's personal copy
Immunoglobulins
P Marnila and H Korhonen, MTT, Agrifood Research Finland, Biotechnology and Food Research, Jokioinen,
Finland
ª 2011 Elsevier Ltd. All rights reserved.
Introduction
Immunoglobulins (Igs), also called antibodies, are pro-
teins present in the milk and colostrum of all lactating
species. The biological function of milk Igs is to give the
offspring immunological protection against microbial
pathogens and toxins and to protect the mammary gland
against infections. To this end, the major mechanisms
provided by milk and colostral Igs are augmenting
phagocytosis and cell-mediated cytotoxicity reactions by
leukocytes, agglutination of bacteria, neutralization of
microbes and toxins, and activation of the complement
system in milk. Igs are divided into classes having differ-
ent physicochemical properties, structures, and biological
activities. The major classes in bovine and human milk are
IgA, IgG, and IgM.
The concentration of different Ig classes in milk and
colostrum varies considerably according to species,
breed, age, stage of lactation, and health status and is
often different from that in blood (Table 1). For exam-
ple, in human milk and colostrum, the IgA class
comprises about 90% and in blood 15–20% of total Ig
whereas in cow the IgG class is dominant in milk,
colostrum, and blood (about 60–70, 80–90, and 90% of
total Ig, respectively).
The transport of Igs from serum to milk is a selective
process, favoring in most species homologous IgG.
Specific receptors are involved in the process, enabling
the characteristic concentration of Ig isotypes in milk
and colostrum. In many species, the absorption of Igs
from intestine is also selective and receptor mediated.
However, in ruminants, for example, in the cow, the
absorption of Igs takes place nonselectively during the
first 12–36 h after birth of the offspring. Ruminant neo-
nates are born virtually without Igs and the colostral Igs
are essential for survival. Thus, in ruminants the colos-
trum contains remarkably higher amounts of Igs than
the mature milk. Mature bovine milk contains only
1–2% of the Igs present in colostrum.
The Ig fraction of milk is used commercially as feed
supplements and replacers of colostrum, mainly for neo-
natal calves and pigs, in order to prevent gastrointestinal
infections. Increasing interest has recently been focused
on the development of colostrum-based Ig products that
contain specific antibodies for the prevention or treat-
ment of infections in humans.
Encyclopedia of Dairy Sciences, Second Edition, 2011, Vol. 3, 807-815
Structure and Functions of
Immunoglobulins
Structure of Immunoglobulin Classes
The Ig classes in milk and colostrum of the cow are
IgG, IgM, and IgA and of human, IgG, IgM, IgA, and
IgD, respectively. The basic structure of all Igs is simi-
lar. They are composed of two identical light chains
(molecular weight of each around 23 kDa) and two
identical heavy chains, each of 53 kDa (Table 2).
There are two types of light chains ( and ), differing
in chain structure but having homologous amino acid
sequences. The light chains contain a constant region
(CL) and variable region (VL). The VL region deter-
mines the immunological specificity. Light chains are
attached to the heavy chains by a disulfide bond and
the two heavy chains are held together by disulfide
bonds near a hinge region. The hinge region gives the
molecule flexibility needed in antibody–antigen interac-
tions. The two identical antigen-binding sites needed in
these interactions are formed by the N-terminal part of
one heavy chain and the variable region of one light
chain. The complete Ig, or ‘antibody’, molecule has a
molecular mass that varies around 160 kDa. The bovine
IgG molecule occurs predominantly in two subclasses:
IgG1 and IgG2 (Figure 1(a)).
Monomeric IgM and IgA have a similar basic structure
to IgG except for the addition of a C-terminal octapeptide
to the heavy chains. IgA occurs as a monomer or dimer,
the latter comprising two IgA molecules joined together
by a J-chain and a secretory component. This complex is
called secretory IgA (sIgA) and has a molecular weight of
about 380 kDa. IgM consists of five subunits, similar to
monomeric IgA, which are linked together in a circular
mode by disulfide bonds and a J-chain; the molecular
weight of pentameric IgM is approximately 900 kDa
(Figure 1(b)).
Human sIgA consists of two identical monomers
joined together by a 15 kDa J-chain and connected to
secretory component, an 83 kDa epithelial glycoprotein.
In human serum, the principal form is a monomer. Both
human serum and milk IgM have a pentameric structure
but, unlike the serum IgM, some of milk IgM is combined
with the secretory component.
In humans the high diversity in structures of variable
chains comprising the antigen-binding sites of Igs is
807
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808 Milk Proteins | Immunoglobulins

Table 1 Concentration of immunoglobulins in serum and mammary secretions of some

species

Concentration
(g l 1)

Species Ig class Serum Colostrum Milk

Cow IgG total 25.0 60 (20–200) 0.47 (0.15–0.8)

(Bos taurus) IgG1 14.0 15–180 0.35 (0.3–0.6)
IgG2 8–11 1–3 0.02–0.12
IgA 0.4 3.5 (1–6) 0.05–0.14
IgM 3.1 5 (3–9) 0.04–1.0
Human IgG 13.5 0.4 (0.16–6.8) 0.04 (0.04–0.054)
IgA 3.5 11.4 (6–40) 0.5 (0.26–1.8)
IgM 1.5 0.3 (0.12–1.9) 0.03 (0.02–0.1)
IgD 30a 0.36a (0.02–
4.1)
Pig IgG 21.5 58.7 3.0
IgA 1.8 10.7 7.7
IgM 1.1 3.2 0.3
Horse IgG total 21.9 113.4 0.39
IgG(T) 8.2 15.2 0.09
IgA 1.5 10.7 0.48
IgM 1.2 5.4 0.03
a
mg l—1
Data extracted from Hurley WL (2003) Immunoglobulins in mammary secretions. In: Fox PF and
McSweeney PLH (eds.) Advanced Dairy Chemistry, Vol. 1: Proteins, 3th edn., pp. 421–447. New York:
Kluwer Academic; Plenum Publishers, Butler JE (1974) Immunoglobulins of the mammary secretions.
In: Larson BL and Smith VR (eds.) Lactation, Vol. III: Nutrition and Biochemistry of Milk/Maintenance,
pp. 217–255. San Diego, CA: Academic Press, Turner 1996, Harzer G and Haschke F (1989) In:
Renner E (ed.) Micronutrients in Milk and Milk Based Food Products, pp. 125–237. Essex, England:
Elsevier Publishers LTD, Brandtzaeg P (1983) The secretory immune system of lactating human
mammary glands compared with other exocrine organs. Annals of the New York Academy of Sciences
409: 353–382 and Prentice A (1995) Regional variations in the composition of human milk. In: Jensen
RG (ed.) Handbook of Milk Composition, pp. 115–221. San Diego, CA: Academic Press, Inc.

Table 2 Properties of bovine immunoglobulins

IgG1 IgG2 IgA sIgA IgM

Physico-chemical
Molecular weight (kDa) 146–163 146–154 160 385–430 900
Heavy chain 56–59 54–59 61–63 62–76
Heavy-chain type 1 2  
Number of H- and L-chains 2 2 2 4 10
Structure Monomer Monomer Monomer Dimer Pentamer
Sw,20 6.3–7.1 S 6.5–7.1 S 6.5–7 S 10.8–11 S 19 S
Carbohydrates (%) 2.8–3.1 2.6–3.0 6–10 10–12
Isoelectric point 5.5–6.8 7.5–8.3
Immunological
Opsonization +++ + 0 0 0
Complement fixation +++ + 0 0 ++
Agglutination + + ++ ++ +++

0 = No activity, + = low activity, ++ = moderate activity, +++ = strong activity. Sw,20 = sedimentation constant in
Svedbergs units (10 13 S).
Data extracted from Butler JE (1974) Immunoglobulins of the mammary secretions. In: Larson BL and Smith VR
(eds.) Lactation, Vol. III: Nutrition and Biochemistry of Milk/Maintenance, pp. 217–255. San Diego, CA: Academic
Press, Butler JE (1998) Immunoglobulin diversity, B-cell and antibody repertoire development in large farm
animals. Revue Scientifique et Technique 17(1): 43–70, Hurley WL (2003) Immunoglobulins in mammary
secretions. In: Fox PF and McSweeney PLH (eds.) Advanced Dairy Chemistry, Vol. 1: Proteins, 3th edn.,
pp. 421–447. New York: Kluwer Academic; Plenum Publishers, Tewari UJ and Mukkur TKS (1975) Isolation and
physico-chemical characterization of bovine serum and colostral immunoglobulin G (IgG) subclasses.
Immunochemistry 12: 925–930, and Zeitlin L, Cone RA, Moench TR, and Whaley KJ (2000) Preventing infectious
disease with passive immunization. Microbes and Infections 2: 701–708.

Encyclopedia of Dairy Sciences, Second Edition, 2011, Vol. 3, 807-815

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Milk Proteins | Immunoglobulins 809

Figure 1 (a) Schematic diagram of a basic immunoglobulin. From Larson BL (1992) Immunoglobulins of the mammary
secretions. In: Fox PF (ed.) Advanced Dairy Chemistry, Vol. 1: Proteins, pp. 231–254. London: Elsevier Science Publishers.
(b) Structure of five classes of immunoglobulins. Adapted from Larson BL (1992) Immunoglobulins of the mammary secretions.
In: Fox PF (ed.) Advanced Dairy Chemistry, Vol. 1: Proteins, pp. 231–254. London: Elsevier Science Publishers and from Butler
JE (1974) In: Larson BL and Smith VR (eds.) Lactation, Vol. III: Nutrition and Biochemistry of Milk/Maintenance, pp. 217–255.
San Diego, CA: Academic Press.

Encyclopedia of Dairy Sciences, Second Edition, 2011, Vol. 3, 807-815

 Author's personal copy
810 Milk Proteins | Immunoglobulins
generated via rearrangements of a large number of diver-
gent Ig gene segments which produce the genes encoding
the variable regions of light and heavy chains, respec-
tively. In contrast, cow and sheep having germline Ig
gene pools of limited diversity employ alternative post-
arrangement mechanisms for antibody diversification.
Cow and sheep generate Ig diversity by non-antigen-
dependent somatic hypermutations. The cow also has an
exceptionally long complementary determining region 3
of the heavy-chain regions of IgM to compensate the
lesser diversity of germline variable heavy-chain genes.
However, the understanding of the processes that form
bovine Ig repertoire is still incomplete.
Functions of Immunoglobulins
All Ig classes bind antigens and, in addition, exhibit one or
more effector functions. While one part of an antibody
(Fab) binds to antigen, other parts (mostly the Fc region)
interact with other elements. In effect, the antibodies
function as flexible adaptors linking various parts of the
cellular and humoral immune system.
The prevention of the adhesion of microbes to sur-
faces like intestinal epithelial linings may be the most
important mechanism of colostral and milk antibodies in
protecting the host. The attachment of a pathogenic
organism to the epithelial lining is the most critical
step in the establishment of infection. The ability of
Igs to agglutinate bacteria reduces the capability of
bacteria to adhere to epithelial surfaces or to tooth
enamel. Normal colostrum and milk are known to con-
tain agglutinating Igs for a large number of pathogenic
and non-pathogenic microorganisms. Igs can inhibit bac-
terial metabolism and reduce the production of harmful
components, such as toxins, by blocking enzymes and
receptors. This blocking may also reduce the pathogen’s
ability to produce structures needed in adherence to
epithelia. Milk Igs, also partially degraded ones, can
benefit the offspring by binding and inactivating toxins
produced by pathogenic microbes and by neutralizing
viruses. In clinical studies, colostral and milk Igs have
been demonstrated to be able to protect the offspring
against a variety of viral infections. Igs also augment the
recognition and phagocytosis of bacteria by leukocytes
(opsonization).
Leukocytes are an integral part of normal milk and
colostrum and are of vital importance in defending the
mammary gland against pathogens. Recent studies have
shown that colostral leukocytes enhance the calf’s
defense against infection. The effector functions include
also binding of Igs to leukocytes or to host tissues. In
blood and tissues, the most important function of Igs is
possibly the activation of complement-mediated bacter-
iolytic reactions, but the significance of this mechanism
in milk remains obscure. Bovine and human colostra
Encyclopedia of Dairy Sciences, Second Edition, 2011, Vol. 3, 807-815
contain an active complement system participating in
the immune defense of the udder (Table 2).
IgM antibodies, although produced in smaller amounts
than IgG, are considerably more efficient than IgG with
regard to most of the above activities, including comple-
ment fixation, opsonization, and agglutination of bacteria.
IgM can prevent the migration of bacteria by binding to
the flagellas used in movement. Bovine IgA, in contrast,
does not fix complement or opsonize bacteria, but agglu-
tinates antigens and neutralizes viruses and bacterial
toxins. The main function of sIgA is to protect the muco-
sal barriers by binding viruses and bacteria, thus
preventing them from attaching to mucosal epithelial
cells. Mucosal IgA also contributes to the antigen pre-
senting. The milk Igs have also been found to act
synergistically with non-specific antimicrobial compo-
nents of milk, such as lactoferrin and lysozyme, as well
as the lactoperoxidase–thiocyanate–hydrogen peroxide
system.
Concentrations of Immunoglobulins in
Milk and Colostrum
Bovine colostrum contains substantially higher concen-
trations of Igs than mature milk. In bovine colostrum, Igs
make up to 70–80% of the total protein content, whereas
in mature milk, the Igs account for only 1–2% of the
protein. The main change from colostrum to normal
milk occurs in the first few days after parturition and
continues at a reduced rate for approximately 5–7 days
(see Colostrum). Table 1 provides data on the concentra-
tions of Igs in serum, colostrum, and milk of selected
mammal species.
In the cow, the major Ig class is IgG. Its concentration
at first milking post-partum ranges from 15 to 180 g l 1, the
mean being approximately 60 g l 1. Thereafter, the IgG
concentration falls sharply to about 1 g l 1 at 12th–14th
milking. The concentration of the various bovine Igs in
serum and lacteal secretions varies according to the breed,
age, health status, and stage of lactation of the animal.
The major IgG subclass of colostrum and milk is IgG1,
which accounts for about 50–80% of total Igs. The aver-
age concentrations of IgG2, IgM, and IgA are relatively
small compared to IgG1 (see Table 1). Bovine colostrum
contains about 1–6 g l 1 of IgA and mature milk
0.05–0.1 g l 1. The concentration of IgM in bovine colos-
trum varies from 3 to 5 g l 1 and from 0.04 to 0.1 g l 1 in
mature milk.
In serum, both IgG subclasses are present at about
equal concentrations (IgG1 11.2 g l 1, IgG2 9.2 g l 1),
while IgA and IgM occur at concentrations of about 0.4
and 3.1 g l 1, respectively (see Table 1).
Cows in their first lactation produce significantly less
(total yield about half or less) IgG than cows in later
 Author's personal copy
lactations due to both a lower IgG concentration and a
smaller volume of colostrum.
In Indian buffalo (Bos bubalus bubalis), the concentra-
tions of Igs in colostrum and milk are of the same order
of magnitude as those of its close relative, the cow
(Bos taurus). The IgG concentration in first colostrum
and in transient milk has been reported to be about 50
and 150% higher, respectively, than in the cow. In
buffalo, IgG1 is the dominant Ig subclass and the content
of IgG2, IgA1, IgA2, and IgM is substantially smaller than
that of IgG1. IgG1 and IgG2 of buffalos and cows have
very similar chemical and biological properties and are
antigenically closely related.
In colostrum of the dromedary camel (Camelus
dromedarius), the major Ig class is IgG but, unlike in
bovines, IgG includes three subclasses IgG1, IgG2, and
IgG3. IgA and IgM are present only in minor amounts.
Camel Ig classes are antigenically very different from
the bovine Igs and have several structural characteristics
different from bovine Igs. In horse and pig colostrum,
IgG is a major and IgA a minor component, but after
transition of colostrum to mature milk, IgA becomes the
receptors
major component (see Table 1). Also, in domestic cat,
dog, and rat colostrum, IgG is the major class but after
receptors for
the transfer of Ig has ceased, IgA becomes dominant in
the milk of these species. In human colostrum and
mature milk, IgA is the dominant Ig class, comprising
over 90% of all Igs, whereas IgG is the most abundant
Ig in blood. The main form of human milk IgA is
dimeric secretory IgA (sIgA), comprising over 90% of
total IgA.
Transfer of Immunoglobulins
Main Pathways
Igs represent the mother’s specific immune response to
the antigens encountered in the past. This immunological
protection is transferred from the mother to the
offspring either in utero or via colostrum or milk. In
the in utero pathway, Igs pass from the mother’s blood
into the fetus across the placenta or yolk sac. In transfer
via colostrum or milk, the Igs, which are derived from
blood or are synthesized in the mammary gland, are
transported through mammary secretory cells into the
colostrum or milk. The ingested Igs are transferred
from the gut across intestinal cells into the blood of
the offspring.
The eutherian mammals can be divided roughly into
three groups according to their Ig transfer pathways.
Species like human and rabbit have only placental
transfer. Human infants are born with serum concentra-
tions of IgG which equal those of their mothers. Most
farm animals (pig, horse, cow, sheep, and goat) have no
Encyclopedia of Dairy Sciences, Second Edition, 2011, Vol. 3, 807-815
Milk Proteins | Immunoglobulins 811
in utero transfer of Igs but an extensive lacteal transport.
The newborns of these species are virtually agammaglo-
bulinemic. Species like dog, mouse, and rat use both
pathways. In marsupials, there is little evidence that
maternal antibody transfer takes place across the
placental barrier but several reports suggest that in
the marsupial pouch, the young depends on passive
immune protection from mother’s milk for a long period
after birth.
Transport Mechanisms
Transfer to milk
The epithelium sheet lining the mammary gland sepa-
rates the milk space from the interstitial space. During the
dry period, the epithelium is permeable to fairly large
molecules, such as Igs, whereas during lactation (except
during mastitis) this lining is impermeable to small mole-
cules. Thus, a vast majority of IgG in bovine colostrum
and milk is derived from blood plasma. The transport by
cellular uptake involves binding of IgGs to Fc
in the basal membranes of the udder. The appearance
and presence of specific high-affinity Fc
IgG1 and lower-affinity receptors for IgG1 and IgG2 on
the mammary gland acinar epithelium corresponds to
the amounts of these subclasses in colostrum and milk at
the time of maximal IgG transport. The Fc receptor-
mediated endocytosis is followed by vesicular transport
and exocytosis at the apical membrane. In contrast to IgA
transport, the receptor does not remain with the IgG in
the milk.
The Igs are produced by B lymphocytes. Some Igs
are carried on the surface of B lymphocytes where they
act as receptors and the others are free in body fluids.
Contact between B lymphocytes and antigen is needed
to cause the B lymphocytes to develop into antibody-
forming and secreting plasma cells. The ruminant
mammary gland contains plasma cells adjacent to the
site of transfer of Igs. These cells produce mainly IgA
but IgM and IgG are also synthesized to a small extent.
The transport of IgA appears to be similar in the mam-
mary secretory cells to that present in other secretory
tissues. Secretory component is synthesized in secretory
cells and moved to the basolateral membrane as a
receptor where it binds dimeric IgA with a J-chain
synthesized in adjacent plasma cells. The complex is
transported through the secretory cell to the apical
membrane where sIgA is secreted by exocytosis and a
segment of the secretory component is cleaved off. IgM
is secreted similarly but a small amount of IgM of blood
origin also appears in milk. IgA in bovine colostrum is
derived partly from intramammary synthesis and partly
from the blood.
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812 Milk Proteins | Immunoglobulins
Transfer from gastrointestinal tract to blood
The passage of ingested Igs across intestinal cells of the
neonate can be specific or non-specific, depending on the
species, and it occurs only for a certain period after birth.
The gut of the suckling calf, pig, or foal is non-selective
and open to the transport of a variety of macromolecules
from 12 up to 36 h after birth, corresponding to the time of
the highest IgG content of colostrum. The main Ig class
of bovine colostrum is IgG1, but the passive transport of
other Ig classes, although in smaller amounts, is also
considered important for the calf. In species which
transfer the Igs both in utero and via colostrum after
birth (e.g., rat and mouse), the absorption of ingested Igs
from the intestine is selective. The Fc
receptors on the IgA may contribute to the prevention of food allergies by
enterocytes of the small intestine mediate selective bind-
ing and transcytosis of IgG until the day of weaning. Also
in the human infant, the colostral Igs are absorbed in small
quantities for a short period of 18–24 h after birth.
Importance of Colostral and Milk
Immunoglobulins to the Offspring
Protection against Microbial Infections
Bovine colostral and milk antibodies represent the cow’s
immune response against a variety of microorganisms
present in the cow’s environment and food. Thus, Igs
contribute to the natural antimicrobial properties of
milk. Maternal milk of farm animals is well known to
protect the newborn offspring against microbial
pathogens.
Receiving colostrum within 12 h after birth is essen-
tial for the survival of newborn calves and piglets.
Newborn calves and piglets which do not receive colos-
trum show a high mortality and poor weight gain during
the first weeks of life. Calves deprived of colostrum or
given colostrum of poor quality promptly suffer severe
long scour episodes. To protect the calf against scouring,
a newborn calf should receive a minimum of 2 l of first
colostrum, containing about 100–120 g Igs, to achieve
a normal serum IgG level of 10 g l 1. Twice as high
mortality rates and higher odds of pneumonia have
been observed in calves with <10 g l 1 serum IgG, with
tripled risk of pneumonia when the dam had clinical
mastitis. The colostrum from the first two milkings is
most important, since the concentration of Igs decreases
steeply after the first milkings (see Milk: Colostrum
and Table 1). A variety of colostral whey- and cheese
whey-based antibody preparations have been used as
feed supplements or colostrum substitutes for neonatal
calves and pigs in order to prevent diarrheal diseases
caused, for example, by rotaviruses and enterotoxigenic
Escherichia coli strains.
In human colostrum, sIgA is predominant, the IgG
being transported via placenta. Dimeric IgA is secreted
Encyclopedia of Dairy Sciences, Second Edition, 2011, Vol. 3, 807-815
by lymphocytes associated with mammary gland epithelia
and is transferred by a polymeric Ig receptor to milk. The
sites of the immune protection by Igs are restricted
mainly to the gastrointestinal tract because the milk Igs
apparently are not absorbed in significant quantities from
the infant gut. Especially IgA antibodies appear to be
particularly important during the first days post-partum,
when the infant’s own mucosal IgA production is defi-
cient. Breast feeding has been shown to reduce the risk of
intestinal infections, morbidity in gastroenteritis and diar-
rhea, otitis media, and pneumonia in suckling neonates.
Breast milk also contains a variety of non-specific defense
factors, which contribute to its antimicrobial effect. Milk
blocking antigen entry at the intestinal surface of the
infant. Low levels of total and specific IgA against food
allergens are proposed to correlate adversely with the risk
of developing allergy to these antigens.
Digestion of Antibodies
IgG, IgM, and IgA without secretory component, when
ingested by humans, are normally degraded by proteases
in the stomach and intestine into small peptides
and amino acids which are subsequently absorbed.
Proteolytic enzymes, pepsin, trypsin, chymotrypsin,
carboxypeptidase, and elastase, initially degrade the anti-
bodies to F(ab9)2, Fab, and Fc fragments. Proteolytic
degradation abolishes their agglutinating properties and
dramatically decreases their functional affinity. However,
F(ab9)2 and Fab fragments from high-affinity antibodies
retain some of the neutralizing and attachment-inhibiting
activity of intact antibodies. The secretory form of milk
IgA is relatively resistant to proteolytic digestive enzymes
in the gastrointestinal tract due to the secretory piece but
can be sensitive to a group of microbial enzymes. About
20–80% of sIgA present in human colostrum passes unde-
graded through the gut of the human infant. In contrast,
the secretory component in IgM is not covalent and does
not provide resistance against enzymatic cleavage.
In addition, bovine milk Igs, which have been sub-
jected to proteolytic conditions of the human intestine,
retain at least partially their immunological activity.
Bovine colostral Igs are quite resistant to gastric acids
but are degraded by proteases and are rather sensitive to
trypsin. However, IgG1 is an exception since it is rather
resistant to trypsin. Bovine colostrum and mastitic milk
contain a compound which inhibits trypsin activity.
A part of bovine colostral IgG1 contains O-glycans which
increase their resistance to pepsin. From 10 to 30% of
orally administered bovine Igs can be recovered from the
stool of human infants and adults. These recovered Igs are
in the form of F(ab9)2, Fab, and Fc fragments. The survival
of IgG in the ileum can be increased by encapsulation.
 Author's personal copy
Utilization of Immunoglobulins
Technological Properties of Immunoglobulins
Milk Igs affect many dairy processes. Igs inhibit or
retard the activity of renneting enzymes of bacterial
origin. In normal milk, this is not noticed due to the
relatively low Ig concentration but, if the Ig concentra-
tion in milk is increased, for example, by adding
colostrum or if the milk is mastitic, renneting may be
retarded. The antimicrobial properties of Igs may
adversely affect fermentation processes. A slower fer-
mentation by dairy starters is noted in colostrum and
mastitic milk, which contain increased amounts of Igs.
Also, high Ig concentrations may adversely affect the
tests for antibiotic based on microbial growth, causing
false positive results.
Igs contribute to cream formation by agglutinating fat
globules, thus accelerating the ascent of cream to the
surface. This phenomenon is attributed primarily to
IgM which have been termed cryoglobulins or ‘cold
agglutinins’. The agglutination property of Igs is, how-
ever, abolished by heat treatment (pasteurization) and by
mechanical agitation.
Among dairy technological processes, the properties
of Igs are affected most by the thermal treatments. In
high-temperature/short-time (HTST) pasteurization
(72  C/15 s) less than 30% of the Ig activity is lost,
whereas ultra high temperature (UHT) treatment
(138  C/4 s) and evaporation processing destroy most
of the specific immune activity of milk due to denatura-
tion of Igs. A rapid heat inactivation of IgG starts at
temperatures higher than 65  C, and at 81  C, as much
as 90% of the virus-neutralizing capacity of Igs is lost in
less than 2 min. On the other hand, heating high-quality
colostrum at 60  C for at least 120 min has no effect on
mean IgG concentration or titer of neutralizing antibo-
dies against bovine viral diarrhea virus type 1. A heat
treatment of 60  C for 30 min has been demonstrated to
destroy viable Mycobacterium bovis, Listeria monocytogenes,
Escherichia coli O157:H7, and Salmonella enteritidis added
to colostrum. Bovine IgG added to UHT-treated milk
has been shown to retain its specific immune activity for
several months. Also, Ig molecules seem to retain their
specific activity well in milk powder, irrespective of the
storage temperature.
Commercial Utilization of Immunoglobulins
Over the last 20 years, considerable progress has
been made in isolation and enrichment techniques
for Igs from bovine colostrum and cheese whey.
Separation technologies, such as membrane filtration
and chromatography, have been used alone or in
combination. Using these methods, the recovery of Igs
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Milk Proteins | Immunoglobulins 813
has varied from 40 to 70% of the level present in the
starting material. Specific chromatographic techniques,
such as immobilized metal chelate chromatography,
immunoaffinity chromatography, and cation exchange
chromatography, have been applied to improve the
yield and purity of Ig preparations.
Commercial whey or colostral antibody preparations
have been used for a long time as feed supplements or
substitutes for farm animals in order to prevent gastro-
intestinal infections. They are used mainly for neonatal
calves and pigs and have proven effective in preventing
diarrheal diseases caused by, for example, rotaviruses and
enterotoxigenic E. coli strains.
If the animal is immunized by a vaccination protocol
against a certain pathogen (or antigenic structure derived
from it) the specific antibody titer of colostrum or milk
may increase up to 100–400 times of that in normal milk
or colostrum. In field experiments, active immunization
of the dam has improved the protection of the offspring
against the pathogens used in the vaccine. Biotechnology
industries have shown increasing interest in possibilities
of raising specific antibodies in bovine colostrum by
hyperimmunizing cows with vaccines made of human
pathogenic microbes or their specific structures like
toxins or other virulence factors. Numerous clinical stu-
dies have been carried in humans with the purpose of
demonstrating the efficacy of hyperimmune colostral pre-
parations in the prevention or treatment of infectious
diseases. Promising results have been reported for the
prevention of infections and also in treatment of diseases
when infection is maintained through recurrent reattach-
ment and reinfection inside the oral cavity or
gastrointestinal tract. Encouraging results have also been
obtained in clinical studies where toxins or inflammatory
compounds are involved and have been neutralized by
specific Igs (see Table 3).
A few immune milk preparations, for example,
against rotavirus, are already on the market. The main
limitation of the clinical use of bovine milk antibodies for
humans is that they originate from a foreign species and
thus can be used only orally against gastrointestinal patho-
gens. To overcome this limitation, trans-chromosomic
cows have been cloned for producing human polyclo-
nal Igs. However, several challenges, for example,
expression of bovine antibody, remain to be overcome
before large-scale production of human Igs for ther-
apeutic purposes can begin. Bovine colostral or milk
products from non-immunized cows that exploit IgG
content have become expensive niche products world-
wide on highly competitive markets for natural
‘wellness’ products.
Currently, the US Food and Drug Administration
(FDA) has accepted the safety of hyperimmune milks on
the basis of clinical studies which show no adverse health
effects from these products.
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814 Milk Proteins | Immunoglobulins

Table 3 Examples of recent clinical human studies on the efficacy of bovine whey or colostral preparations

Microorganism used in
immunization Target disease Treatment regimen Efficacy References

Five or one strains of
Escherichia coli
No immunization
No immunization
No immunization
Clostridium difficile toxin
and C. difficile whole
cells
C. difficile whole cells
C. difficile toxin and
C. difficile whole cells
C. difficile toxin and
C. difficile whole cells
Diarrhea
Upper respiratory tract
infections
Mild hypercholesterolemia
HIV-associated diarrhea
C. difficile diarrhea
C. difficile diarrhea
C. difficile diarrhea
C. difficile diarrhea
Once per day orally 0.5 g Lower incidence of diarrhea Tawfeek
of IgG per kg of body and shorter duration of et al.
weight, follow-up diarrhea episodes (2003)
period for 6 months
60 g of colostral protein Reduced significantly Brinkworth
daily for 8 weeks incidence of self-estimated and
symptoms of respiratory Buckley
infections but no difference in (2003)
duration
Orally 5 g of blood Both total cholesterol and LDL Earnest
derived IgG daily for 3 levels decreased from et al.
or 6 weeks baselines (2005)
Orally for 4 weeks Reduced diarrhea and Florén et al.
increase in body weight, (2006)
decrease in fatigue
Orally for 2 weeks as C. difficile toxins eradicated Van Dissel
supportive treatment from 15 of 16 patients and no et al.
after antibiotic relapses in any patient during (2005)
treatment 11-month follow-up period
Orally 1.6 g of Ig in At day 14 after starting 89% of Mattila
200 ml twice per day patients had responded et al.
for 2 weeks positively to the treatment (2008)
Orally 5 g of whey Clinical evidence for the safety Young et al.
powder for 2 weeks as for use in C. difficile patients (2007)
supportive treatment
after antibiotics
Orally 5 g of whey Relapse in 10% of Ig-treated Numan
powder 3 times daily patients whereas relapse in et al.
for 2 weeks for 20–25% of control group (2007)
supportive treatment
after antibiotics

Brinkworth GD and Buckley JD (2003) Concentrated bovine colostrum protein supplementation reduces the incidence of self-reported symptoms of
upper respiratory tract infection in adult males. European Journal of Nutrition 42(4): 228–232.
Earnest CP, Jordan AN, Safir M, Weaver E, and Church TS (2005) Cholesterol-lowering effects of bovine serum immunoglobulin in participants with
mild hypercholesterolemia. American Journal of Clinical Nutrition 81(4): 792–798.
Florén CH, Chinenye S, Elfstrand L, Hagman C, and Ihse I (2006) ColoPlus, a new product based on bovine colostrum, alleviates HIV-associated
diarrhoea. Scandinavian Journal of Gastroenterology 41(6): 682–686.
Mattila E, Anttila VJ, Broas M, et al. (2008) A randomized, double-blind study comparing Clostridium difficile immune whey and metronidazole for
recurrent Clostridium difficile-associated diarrhoea: efficacy and safety data of a prematurely interrupted trial. Scandinavian Journal of Infectious
Diseases 40(9): 702–708.
Numan SC, Veldkamp P, Kuijper EJ, van den Berg RJ, and van Dissel JT (2007) Clostridium difficile-associated diarrhoea: Bovine anti-Clostridium
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