Lch s

Bi 5

Bnh u ma - Smallpox

Sn xut vaccin
Yu cu :

Xc p Pharaoh Ramesses V ca Ai Cp,

1157 BC

1. Phn bit c cc loi vaccin.

China/India, Nht
Cuc Thp t chinh

Ty u: t l cht 25%

2. Nu qui trnh c bn sn xut vaccin vi khun

Ring TK XX: 300 triu ngi cht

3. Nu qui trnh c bn sn xut vaccin virus

4. Trnh by mt s t cht min dch thng dng

Lch s

Lch s

Bnh u ma

Bnh u ma
Vaccination

Chng u

Jenner 1796 : u b/u ln

China/India 1% so vi 25% cht

1800s Tim phng ph cp cho tr em

Min dch ko di

1930s Ca t nhin cui cng Anh

UK: 1700s

1940s Ca t nhin cui cng M

China 1950

1958 chng trnh WHO

Pakistan/Afghanistan/Ethiopia
1970

10/1977: Ca cui cng (Somalia)

Edward Jenner (1749-l823)

nh ngha
Vaccin l ch phm cha khng nguyn ca
mt hoc nhiu loi vi sinh vt, dng
gy min dch ch ng d phng

Vaccin
Thnh cng trong qu kh
Trin vng cho tng lai

Vaccin l tng
 Sinh min dch
Bt chc nhim t nhin
 Cho s bo v ko di
Khong cch gia cc liu xa
 Khng gy tc dng ph nghim trng
 n nh
5

Phn loi vaccin

Phn loi vaccin truyn thng
Vaccin bt hot (vaccin cht)
Vaccin sng gim hot lc
Vaccin c t
Vaccin di n v

Phn loi vaccin

Mt s vaccin mi
Vaccin idiotyp
Vaccin lin hp
Vaccin vector ti t hp
Vaccin DNA
Vaccin tng hp
7

Phn lo
i vaccin

Phn lo
i vaccin

Vaccin bt hot (vaccin cht)

Vaccin sng gim hot lc

cha cc vi sinh vt sng c lm suy yu c lc hoc vi
sinh vt gn ging nhng t nguy him hn( BCG)

Cha cc vi sinh vt c c lm cht bng cc cht ha

hc hoc nhit , do khng cn sinh si trong ch th
c na

u:

Kch thch khng nguyn lin tc lu sn xut t

bo nh  to p ng min dch bn

u: ni chung an ton, nhng khng phi l tuyt i

Cc ni c t b mt trn vaccin ho g bt hot i khi
cm ng cc p ng DTH v virus cm c gn vi cc
phn ng tng t

Nhc
C th chuyn li thnh dng gy bnh

Nhc:
. Thng t hiu qu hn vaccin sng gim c lc
. Ch c kh nng cm ng min dch th dch v mm bnh
cht khng th i vo t bo ch
khng tc dng nu l
nhim ni bo
. Cn nhiu liu hn cho bo v lu di

V d vaccin chng li cm, t, ho g, di, dch hch, vim

gan A v vaccin bi lit Salk

V d
Vaccin chng li st vng da, si, rubella, quai b v
vaccin bi lit Sabin, BCG

10

Phn lo
i vaccin

Phn lo
i vaccin

Vaccin di n v

Vaccin c t

Cha mt phn nh ca tc nhn gy bnh sn xut t vi sinh

vt an ton hoc t bo nui cy gy ra p ng min dch.
Bng cch ny, c th trnh c cc c t mnh, hoc loi
b c cc vt liu m h hoc p ng min dch ln t

cha cc c t bt hot khng cn gy bnh

V d
cc vaccin c t un vn v bch hu,
vaccin c t Crotalis atrox chng li rn chung
cn dng cho ch

u: . t tc dng ph hn vaccin c in
. An ton hn

V d
vaccin Haemophilus influenza type b,
vaccin chng li HBV ch cha cc protein b mt ca virus
vaccin tiu phn ging virus chng li HPV cha protein
capsid chnh ca virus
11

12

Phn lo
i vaccin

Phn lo
i vaccin

Vaccin khng-idiotyp

Mt s vaccin mi

cha khng th khng idiotyp

Vaccin idiotyp
Vaccin lin hp

KT
2

Vaccin vector ti t hp
Vaccin DNA

KN

Vaccin tng hp

KT1

Anti-idiotype

Iditotype

KT 3
KT 3 = KT khng Iditotype

13

14

Phn lo
i vaccin

Phn lo
i vaccin

Vaccin khng-idiotyp

Vaccin idiotyp (tt)

Virus
Khng th
khngidiotyp

epitop
Khng th

Khng th c v
tr gn ca epitop

khng th

15

To khng th
khng li khng
th idiotyp
Khng th khngidiotyp bt chc
epitop

16

Phn lo
i vaccin

Vaccin idiotyp (tt)

Vaccin lin hp

Dng khng th khng-idiotyp lm vaccin

Vaccin polysaccharide

Dng nh
vaccin

Khng th
khng idiotype

Gn v trung
ha virus

Khng th cho khng

th khng-idiotyp

V ngoi polysaccharide l KN khng ph thuc Tcell, to min dch yu tr nh, khng to hoc t tr
nh min dch, nh hng nhc li km
 lin kt cc ny vi protein (c t)
KN protein
. Vaccin chng vim phi do ph cu cha 23 loi
polysaccharide t nang ca 23 chng St. pneumoniae

Khng th
khng-khng-idiotyp
Khng th
khng-khng-idiotyp

Phn lo
i vaccin

. Vaccin Haemophilus influenzae type B

Khng th
khng-khng-idiotyp
17

Phn lo
i vaccin

Vaccin vector ti t hp
L kt hp sinh l ca mt vi sinh vt v DNA ca
vi sinh vt khc, chng li cc bnh c qu trnh
nhim phc tp.

18

Vaccin ti t hp
dng gi mang l virus u b

Phn lo
i vaccin

Virus u b thuc h Orthopoxvirus

Sinh sn trong nhiu vt ch

V d

Virus ADN gm 150-200 gen, ADN xon kp c

187.000 cp nucleotid bao bc bi v lipid

Vaccin phng mt s bnh do virus c lm bng

cch ti t hp bin chng virus u b vi cc
gen v hi ca virus gy bnh,

Lo
i tr mt phn h gen ca virus u b 

khng cn c tnh gy bnh no n
a

Gi mang Mycobacterium bovis (chng BCG)

19

20

Vaccin ti t hp
dng gi mang l virus u b

Phn lo
i vaccin

Vaccin ti t hp
dng gi mang l virus u b

Phn lo
i vaccin

To virus ti t hp
u im

ADN m ha cc khng nguyn virus gy bnh c

phn lp v bin i gen th hin c trong ADN
ca virus u b  Ghp vi ADN ca virus u b

Vaccin virus u b c tc dng min dch

ko di do n nh v mt di truyn

 gen m ha ngng kt hng cu t ca virus cm

C th dng bng nhiu ng 

chng trn qui m ln cc gia sc gia cm,
ng vt hoang di

 khng nguyn b mt ca virus vim gan B

 glycoprotein ca virus herpes thng thng, virus di,
virus Epstein-Barr, virus HIV, virus gy vim ng tiu
ha d ly..

Khng kh bo qun vaccin

Lm vaccin a gi gm 20 loi KN virus khc nhau

21

22

Phn lo
i vaccin

Phn lo
i vaccin

Vaccin ADN

Vaccin ADN

1796 Jenner: virus ng vt

1800s Pasteur: virus suy yu
1996 vaccin ADN

Gene
cho KN

plasmid

T bo c

The third vaccine revolution

T bo c biu hin
protein to khng th
23

Cytotoxic T Lymphocyte (CTL)

p ng CTL

24

Phn lo
i vaccin

Vaccin ADN

Phn lo
i vaccin

Vaccin ADN
C th sn xut vaccin ph rng nu dng hn hp
plasmid m ha cho nhiu on protein t mt hay
nhiu loi virus

Plasmid d sn xut lng ln

ADN rt n nh
ADN khng vi nhit  tin bo qun v vn
chuyn

Plasmid khng sao chp v ch m ha cc protein

quan tm

Trnh t ADN d bin i trong phng th nghim 

Ta c th i ph vi cc thay i ca tc nhn nhim

Khng c thnh phn protein nn s khng c p

ng min dch chng li vector

Bng cch dng plasmid trong vaccin m ha cho

tng hp khng nguyn, protein khng nguyn c
sn xut c bin i hu dch m theo cng cch
vi protein ca virus chng li s phng v khi i vo

To khng nguyn tt hn l protein tinh khit

Cch trnh din ny ca khng nguyn  p ng

CTL trc tip chng li bt c khng nguyn no ca
mm bnh. p ng CTL cng phng v cc bnh
gy bi cc tc nhn ni bo bt buc (v d M.
tuberculosis)

25

26

Phn lo
i vaccin

Vaccin ADN

Phn lo
i vaccin

Vaccin ADN

Cc vn c th:

Cc vaccin ADN gy sinh si t bo nhim virus

S tch hp plasmid vo b gen ch c th gy

t bin chn

Cho:
p ng min dch ph rng

Cm ng p ng t min (v d cc khng th
khng-ADN mm bnh)

p ng CTL ko di
u im ca vaccin ADN cho cm:

Cm ng dung np min dch (v d ni biu hin

khng nguyn trong ch th c th c p ng
khng c hiu vi khng nguyn )

p ng CTL c th chng li protein ni sinh

chut vaccin ADN nucleoprotein c hiu qu
chng li nhiu virus c hemagglutinin khc nhau

27

28

Phn loi vaccin

Phn lo
i vaccin

Vaccin tng hp

Mt s tn phn loi vaccin khc

Cha cc peptid, cc carbohydrat hoc cc khng

nguyn tng hp phn chnh hoc ton phn

Vaccin hp ph: Vaccin DPT c hp ph trn gi

gel nhm hydroxyt.
Vaccin khc chng: Vaccin c tc dng chng li vi
sinh vt khc vi loi dng ch n. V d: vaccin t u
b phng u ngi.

Vaccin peptid tng hp vi khun

Trung ha c t
Vaccin peptid tng hp c t bch hu v t

Vaccin a gi: Vaccin cha cc khng nguyn bo v

ca nhiu chng thuc v mt loi vi khun gy bnh. V d:
TAB.

Vaccin peptid tng hp virus

Cc vng khng bin i
Cc vaccin peptid tng hp cho cc protein v
glycoprotein HIV ang c pht trin

Vaccin hn hp: Vaccin gm nhiu khng nguyn ca

cc loi vi sinh vt khc nhau, phng nhiu bnh nhim
trng. V d: DPT, Priorix.

Vaccin peptid tng hp k sinh trng

vaccin peptid tng hp cho cc epitop bo t
k sinh trng st rt

29

30

Quy trnh sn xut vaccin in hnh

PHN CHNH BO CH

Phng php
sn xut vaccin

NG NG

NG GI

Khng nguyn 1

Khng nguyn 2

Khng nguyn 3

T cht

31

32

Phng php sn xut vaccin

Phng php sn xut vaccin

1. H thng ging
. Phn lp vi sinh vt thch thch hp

1. H thng ging

. t cc nhim trng ngi

. thao tc v chn lc thm

2. Sn xut vi khun v cc thnh phn vi khun

ca vaccin vi khun

. Lm l ging (seed lot)

3. Sn xut virus v cc thnh phn virus ca

vaccin virus

. nhn vi sinh vt thch thch hp ln

. phn vo ng
. bo qun -700C hoc ng kh

4. Phi trn
5. ng ng v lm kh

. Kim tra an ton v hiu qu th lm sng. Khi

cc kt qu th lm sng t cng nhn, l ging
c dng thng qui cho sn xut vaccin

6. Kim sot cht lng

33

34

Phng php sn xut vaccin

Phng php sn xut vaccin

Ln men vi khun
2. Sn xut vi khun v cc thnh phn
vi khun ca vaccin vi khun

- Hot ha ging vi khun t - 700C /ng kh

Ln men

- Cy chuyn mt hoc vi ln vo mi trng nhn

ging  lng

Thu vi khun

- Cy chuyn ging vo mi trng ln men

khuy lin tc, theo di v iu chnh pH v th
oxy ha kh ca mi trng sut qu trnh nui
- Thi gian: 1 ngy - 2 tun
 Thu hoch

35

36

Phng php sn xut vaccin

Phng php sn xut vaccin

Qu trnh thu vi khun

Thu vi khu n l tc nhn gy bnh, nguy him
Sn phm ln men:

1 Git cht

t bo vi khun,
cc sn phm chuyn ha v
mi trng cn kit

2 Tch
3 Phn on
4 Kh c t

Vaccin sng gim hot lc

. tch vi khun khng c ra
. ha vo dung mi thch hp, c th ng kh

5 Hp ph
6 Kt hp

37

38

Phng php sn xut vaccin

Phng php sn xut vaccin

Thu vi khu n l tc nhn gy bnh, nguy him

Thu vi khu n l tc nhn gy bnh, nguy him

1. Git cht

3. Phn on

Dng nhit v cht ty u.

. Chit cc thnh phn t t bo hoc t mi trng

nui vi khun

Bordetella pertusis lm vaccin ho g - dng un

nng /hoc formalin

. Tinh ch bng dung mi thch hp v kt ta

. Lm kh thnh bt, bo qun v hp nht vo vaccin

Vibrio cholerae trong vaccin t v Salmonella typhi

trong vaccin thng hn - dng phenol

4. Kh c t

2. Tch

chuyn c t ca vi khun thnh c t khng hi

dng ly tm

. dng Formalin kh c t ca C. diphthe v C. tetani

Thu t bo v ha vo hn hp mui hoc

. Tin hnh trn dch nui cy trong ni ln men hoc

trn c t tinh khit sau khi phn on

Thu dch lng, b t bo

39

40

10

Phng php sn xut vaccin

Thu vi khu n l tc nhn gy bnh, nguy him

Phng php sn xut vaccin

3. Sn xut virus v cc thnh phn virus

ca vaccin virus

5. Hp ph
Cht hp ph: t cht khong, nhm hydroxid, nhm
phosphat v canxi phosphat  tng tnh khng
nguyn v gim c tnh ti ch v h thng ca
vaccin.

Nui virus
Thu virus

Vaccin bch hu v vaccin bch hu/un vn/ho g

6 Kt hp
. tng cm ng p ng min dch
. Polysaccharide nang ca H. influenzae Type b + c
t bch hu v un vn + protein mng ngoi ca
Neisseria meningitidis
41

42

Phng php sn xut vaccin

Phng php sn xut vaccin

Nui virus

Thu virus

. Nui virus trong phi g, vt hoc nui cy m

- Chn m thch hp cho virus
- Cn tuyt i v trng trong qu trnh nui cy,
kim nghim ngt ng vt tuyt i KHNG
c nhim virus l vo phi

Lc v thu dch c cha virus

. Trng g c phi
Virus cm tch t trong dch ti niu ca trng
Virus st vng da tch t trong h thng thn kinh
ca phi

Vaccin cm: ly tm dch ti niu  huyn trc virus

tinh khit mt phn v m c  x l vi ete
hoc vi tc nhn ph v khc ct virus thnh
cc thnh phn ca n nu mun c vaccin virion
hoc khng nguyn b mt.
Vaccin st vng da: ng ha phi g trong nc
thnh bt nhuyn cha virus  ly tm kt ta hu
ht mnh v ca phi v virus st vng da

Nui cy chm
43

44

11

Phng php sn xut vaccin

Thu virus

6. Kim sot cht lng

. Nhm m bo v hiu qu v an ton ca
mi l sn phm

. Khng c giai on bt hot v hu ht vaccin lm t

nui cy t bo cha virus sng suy yu
Ngoi tr 2 trng hp
Vaccin virus bi lit bt hat c bt hot vi
formalin long hoc 3-propiolactone  hp ph v
ra
Vaccin di c bt hot vi 3-propiolactone 
siu lc

. Thc hin theo ba cch: (1) kim sot trong qu

trnh; (2) kim sot sn phm cui v (3) yu cu
ph hp cho mi nguyn liu sn xut ban u,
trung gian, sn phm cui v cc phng php
x l. Cc kt qu kim tra kim sot cht lng
lun c ghi chi tit.
Kim tra an ton gm cc kim tra tng qut
l: (1) v trng, (2) Khng c c t bt
thng, (3) S hin din ca nhm v calci, (4)
Formalin t do v (5) Nng phenol

. Khi hon tt, khi nguyn liu c bo qun nhit

-700C n khi trn thnh vaccin cui

45

Phng php sn xut vaccin

V d

46

Sn xut vaccin vi khun thng hn

Nhn ging

Nui Salmonella typhi trn thch / 18 gi

Ha 5x108 vi khun /1ml NaCl ng trng

Nui cy

Mi trng lng casein

Cy ging 5%-10% V
Nui cy lin tc c h thng khuy 180-300 vng/1 pht
Sc kh v trng 1lt/1pht; Thm dung dch glucose 40%
pH 7,6-7,8
10-12h

Lm bt hot

6x1010 vi khun/ 1ml

Trn sinh khi vi EtOH 960 ln 1- 1:4 , ln 2- 1:10

Kim tra tinh khit v p ng min dch

Bo ch a dung dch v im ng in bng NaCl c cha 0,25%
phenol, 1ml vaccin cn cha 5x109 vi khun.
ng ng v ng kh - 40
- 500C
ng gi

47

48

12

Sn xut khng nguyn Vi (v) tinh khit t

vi khun thng hn Salmonella typhi
Nui cy

Vaccin nga un vn

Vi = virulence

Lm kh sinh khi bng cn

Clostridium tetani

Mi trng lng casein + dch men bia thy phn

Nui cy k kh
 Thu exotoxin
hp ph vi Al(OH)3
 Lm bt hot
formalin
exotoxcin
anatocin
 Tinh khit ha
hp ph li vi Al(OH)3
 Kim tra tinh khit v p ng min dch
 Bo ch
1 ml vaccin cha 20 n v ngng kt, pH 6,8-7,6

Chit bng nc ct (3 ln)

Ta bng cn
Ln 1: cn + NaCl bo ha
Ln 2: cn pH 3,5 3,8
Ln 3: cn 46-52%
ng kh
Bo ch
Ha khng nguyn Vi trong NaCl ng trng 40 g/ml
Khng nguyn Vi dng  d
ng mono vaccin ho c
chung vi vaccin vi khu n ho c phi hp vi antigen O (= mng)

 Ging

 Nui cy

49

Sn xut vaccin chng vim gan B

50

Sn xut vaccin chng vim gan B

Nguyn nhn: HBV, truyn qua ng tim chch, tnh dc

M: 200.000 ngi nhim/nm, trong 20.000 ngi mang
mm bnh v 1/5 s ny cht do x gan, 1/20 b ung th gan
1963: pht hin khng th HBV trong mu bnh nhn a chy
mu
1968: HBsAg
nui HBV trn t bo gan - KH

Nguyn tc: trch khng nguyn b mt HBsAg ca virus t mu

ngi b nhim virus hepatitis B mn tnh
Khng dng phng php nui cy m c y l virus i gan
m t bo gan nguyn thy l
i kh nui cy
Cc bc chnh
Mu ngi nhim vim gan siu vi B mn tnh
Bt hot bng permalin, To v hydroxyt nhm hoc
pepsin, urease v formalin
Tinh ch khng nguyn

1970: huyt thanh ngi nhim HBV mn

Th c tnh v p ng min dch

Th c tnh trn tinh tinh / 65 tun cho mi l vaccin
Th hot tnh khng nguyn ca vaccin trn chut lang
v chut nht
51

LU NHT, T TIN NHT

52

13

Sn xut vaccin chng vim gan B

ti t hp

Sn xut vaccin chng vim gan B

ti t hp

i t t bo ng vt
a gen m ha khng nguyn vo t bo ng
vt c v bng cc vector nh adenovirus,
virus SV40 hoc virus gy u v b

i t t bo vi khu n v nm men

phn lp gen m ha HBsAg
a gen ny vo E. coli hoc S. cerevisae
sn xut HBsAg bng nui cy vi khun hoc nm men
tch khng nguyn t t bo 1-3 g/1ml dch nui cy

u im
Cc virus ny c th sao chp c lp trong t bo
nh nhng plasmid vi khun
Cc t bo ng vt c v c kh nng khuych i
ln cc khng nguyn: 1 ml dch nui cy cho 24 g
khng nguyn
T l glycozyl ha cao, d tch khng nguyn t dch
v khng nguyn c sn xut lin tc

53

54

Vaccin cm

Sn xut vaccin chng vim gan B

San xuat vaccin chong HBV: so sanh pp truyen thong va tth
Pp truyn thng
huyt thanh ngi mang HBV
Bt hot (formaldehyde)
Tinh ch khng nguyn
Th c tnh v p ng min dch
(tinh tinh/ 65 tun cho mi l)
Th hot tnh (chut lang v chut nht)
40 lt


1 liu vc-xin
Pp mi 40 lt dch nui cy S. cereviseae ti t hp mang gen m
ha protein S t HBV
nhiu liu vc-xin

55

S thay i KN virus cm A
1890 H2N8
1900 H3N8
1918 H1N1
1957 H2N2 (cm chu )
1968 H3N2 (cm Hong Kong)
1977 H1N1
1987 H3N2
Cm A c 15 hemagglutinin (H1 --> H15) v
9 neuraminidase (N1, N2, ...)

56

14

Pht trin vaccin cm

Vaccin cm g H5N1

Nhm H glycoprotein b mt m virus cm dng gn vo t

bo ch th
Trivalent vaccine - t hp cc protein t 3 chng ht ging
1 hoc 2 trong 3 c thay i mi nm
Mi chng l t hp trnh t tng trng tt trong trng
vi trnh t KN t virus cm
Virus mc tt trong trng + Virus c ch KN mong mun
Cng nhim vo trng g c
phi
8 mu gen ti t hp
Chn lc
Virus c H & N mong mun+ mu gen mc tt trong trng

Vn
virus git trng g dng nui chng ht ging qu sm
H5 hemagglutinin c trnh t c c tnh & git trng g

Dng di truyn ngc to chng ht ging
cDNAs 8 mu hi li trong t bo nui cy
Bin i mt phn trnh t ny
Ht ging nui c trong trng
Birmingham adenovirus biu th hemagglutinin trong dng
t bo ngi PER.C6
(th Pha 1)
Meriden, Con-base Protein Sciences Corp
FluBlOk
rHA vect baculovirus trong t bo cn trng
(th Pha 2 ti Univ. Rochester)

57

58

Ti t hp khi cng nhim hai virus

(Recombination by independent assortment during dual 59
infection)

Ti t hp khi cng nhim hai virus

(Recombination by break-rejoin of incompletely linked
60
genes)

15

Vaccin Bi lit

Ti t hp khi cng nhim hai virus

(Recombination by copy-choice of incompletely linked
genes. The genetic interaction of certain RNA viruses can
61
result in copy-choice recombination)

62

Bai llet

1789: c m t ln u bi Micheal Underwood

1843: bng pht dch u tin c m t M
1921: dch bng pht TT Roosevelt
1952: 21.000 ca bi lit M
Poliovirus: Enterovirus, c 3 tp huyt thanh 1, 2, 3;
Nhanh chng v hot bi nhit, formaldehyd,
chlorine, tia UV

Bai llet

Vaccin virus bi lit

1955 Vaccin v hot (inactivated vaccine)
1961 OPV mt loi virus tp 1 v tp 2
OPV mt loi virus tp 3
OPV ba loi virus
1987 IPV tng cng hot lc (IPV)

2 loi vaccin bi lit:

I- Jonas Salk pht minh, cha virus bi lit cht, tim
II- Albert Sabin virus sng suy yu, ung,
nm 2000 b ngng, chi dng ni nguy c cao

63

64

16

Bai llet

Bai llet

Vaccin bi lit v hot

Cha 3 tp huyt thanh ca virus
Tng trng trn t bo thn kh (Vero)
V hot bng formaldehyde
Cha 2-phenoxyethanol, neomycin, streptomycin,
polymyxin B
Vaccin bi lit ung
Cha 3 tp huyt thanh ca virus
Tng trng trn t bo thn kh (Vero)
Cha neomycin v streptomycin
Bi tit trong phn ngi trong 6 tun sau khi
chng
65

66

Bai llet

Bai llet

1988: khi ng chng trnh thanh ton  2005

Gim bi lit t hng trm ngn ca/nm 1000 ca/nm
8/7/2005: bng pht dch Angola v Indonesia
Vit Nam
Chng trnh thanh ton SBL thuc chng trnh
tim chng m rng
1996 2 ca
2000 c cng nhn hon ton thanh ton
(Indo 1995)

67

68

17