Biotechnological aspects

of Product Development
Rana Muhammad Awais Khan
PHM-753
(Formulation & Product Development)

M.Phil. 2nd
Department of Pharmacy
Quaid-i-Azam University Islamabad
 Quality control of Biotech
Products
Issues in Biotechnology Products
Ahsin Iqbal
PHM-753
(Formulation & Product Development)

M.Phil. 1st
Department of Pharmacy
Quaid-i-Azam University Islamabad
Quality Control of Sterile Products

There are three general areas of sterile Quality control which includes:
1. Incoming stock Control
2. In process Quality Control
3. Finished Product Control

1. INCOMING STOCK CONTROL

For sterile product, incoming stock control include routine confirmation of color, odor, crystalline state, solubility, identification, melting point,
loss on drying and specific gravity of all ingredients as well as special evaluation such as
• Pyrogen testing for water for injection • Glass test on container • Identity of rubber closure etc.

2. IN-PROCESS CONTROL
In-process control for the manufacturing of sterile product includes; tests, readings and observation made throughout all the manufacturing
process of product which involves;
• Laminar Air Flow: It is an environment within which airflows in a confined area with uniform velocity along parallel flow lines with
minimum swirls of 99.99% efficiency.
• Class 10,000 Clean: The area in which particles count in the air is not more than 10,000/feet3 of size 0.5micron or larger.
Class 1000 Clean: The area in which particles count in the air is not more than 1000/feet3 of size 0.5micron or larger.
Class 100 Clean: The area in which particles count in the air is not more than 100/feet3 of size of 0.5micron or larger.

Other tests: Other things which should be seen during in process control of biotechnological products are;
 conductivity measurement during distillation of water for injections
 confirmation of volume of fill,
 confirming the label,
 quantity,
 rubber closures and vials.

3. FINISHED PRODUCT CONTROL

This place includes all the final assays and tests to which finished product is subjected. It includes following tests;
•Leaker Test • Clarity Test • Pyrogen Test • Sterility Test • Assay of Active Ingredients
Leaker test:
 Leaker test is intended to detect incompletely sealed ampoules so that they may be discarded. Ampoules are intended to provide a
hermetically sealed container for a single dose of product, thereby preventing any interchange between the contents of the sealed ampoule
and its environment.
There are two types of sealing; 1. Tip sealing 2. Pull sealing
 Tip sealing ampoules are more likely to be incompletely sealed than those that have been pull sealed. In addition, some time cracks may
appear around the seal or at the base of the ampoule as a result of improper handling.
Elimination
1. By Heating:
At 1800C for 4 hours At 2000C for 1 hour
At 2500C for 40-45 minutes At 6500C for 1 minute
2. By Distillation 3. By Filtration 4. By Adsorption

Q.C Tests for Pyrogen

Pyrogen can be tested in two ways i.e.
1. IN-VITRO TESTING
• It is also known as “LAL” test. LAL stands for stimulus amebocyte lysate
• It is based on the principle of turbidity formation of pyrogen preparation in the presence of LAL (Turbidity will occur if pyrogen is present
in solution by LAL).
• This enzyme reacts with gram negative bacterial endotoxin (pyrogen) so that assay mixture increased in viscosity and opacity until an
opaque gel is formed.
PROCEDURE
• Mix equal volume of test solution and LAL reagent (0.05-0.2ml) in thoroughly cleaned, disposable and dry heat sterilized test tubes.
• Incubate the mixture at a temperature of 37°C for 1 hour. Care should be taken that during incubation, test tube must be undisturbed because
shaking may cause irreversible removal of turbidity leading to false results.
• Observe the test tubes and examine for turbidity formation to reach the end point.
2. IN-VIVO TESTING
• This is a biological test which employs rabbit as a test animal. It rises the body temperature evoked in rabbits by IV administration of sterile
injection of sample been examined.
• Thermometer and Electrical Devices are used to note the temperature.
• Thermometer must be inserted into the rectum of the rabbit at a depth of 5.2cm according to B.P 7.2cm according to U.S.P In case of
electrical devices, these are inserted into the rectum at least 30-90 minutes before injection.

CLARITY TEST
In its subjective sense, it means to detect or spot with naked eye any particulate matter in ampoule or otherwise.
Clarity is relative term, the meaning of which is markedly affected by the factors like Visual activity of the observer, Meaningful interpretation
of what is seen, Degree of critical thought, preparatory to the inspection (moving particle can easily be determined), Physical alertness.
Particulate Matter:
It includes dust, lead, carbon particle, rubber, glass particles, fibers etc. Upon injection they may produce pathological granulomas. So, this
particulate matter must be removed and parenteral must be free from these particles of 50 meau can be detected by naked under suitable
condition of visibility. Each particle has approximately 600 meau diameter. So, easily be detected by naked eye.
Procedure:
Clarity test is performed by the evaluation of sample under suitable condition of visibility by the human inspection in the presence of white and
black background, while the contents of the container are set in swirling mode because it is stated that a moving particle can easily be detected
as compared to one that is stationary.
 ISSUES IN BIOTECHNOLOGICAL PRODUCTS

A. Building capacity for developing and managing biotechnology:

Application of biotechnology to meet the needs of developing countries requires the creation of an infrastructure for the
transfer of relevant technologies, development of institutions with the capacity to adopt and develop the know-how required
for successful application of biotechnology. This includes building capacity to understand their own ecosystems and to select,
acquire, manage and further develop those biotechnologies that are most appropriate to national needs. Clearly, such efforts
require investing in science and technology education and research.

B. Biosafety and bioethics: capacity for risk assessment:

Biosafety is concerned with the potentially adverse impacts of biotechnology on human, animal and plant health, and the
environment. Biotechnology also gives rise to socio-economic and ethical concerns, some of which have been described here.
Physical risk and uncertainty are technical issues, and policies and regulatory regimes intended to manage these risks will
depend largely on scientific capacity, including human expertise and well-equipped laboratories. This capacity simply does
not exist in many developing countries at present.

C. Building awareness of biotechnology:

Some of the applications of biotechnology described earlier have potentially serious implications for socio-economic welfare,
and ethical and moral well-being. If biotechnology is to be used to provide benefits to a country’s population, then political
support, as well as public awareness and acceptance of new technologies are essential. There is a wide range of potential
applications, and decisions have to be made concerning the choice of technologies, according to national needs. The public has a constructive
role to play in helping to make these choices, but in most countries, including industrialized countries, public awareness and knowledge about
biotechnology are insufficient for ordinary people to have an effective and qualified voice in biotechnology development. Building public
awareness and disseminating qualified and balanced information about biotechnology is a critical issue in most countries.

D. Accessing biotechnology: intellectual property rights:

Many of the new products and processes associated with biotechnology have been developed in the private sector, and this has led to concerns
that proprietary rights to these technologies might mean that many developing countries will be unable to access them. Another issue is that it is
felt by many that ownership rights of genes and other living matter, as intellectual property, is not morally acceptable. Furthermore, the
patenting of gene sequences and biotechnology techniques with broad applications means that developing countries in particular may be
excluded from affordable access to technologies that they urgently need. Against this, innovating organizations argue that without the limited
monopoly rights to profit from their new products and processes that are conferred by intellectual property tights (IPRs), there is no incentive to
invest in research and development. Moreover, some argue that where IPRs cannot be adequately protected, this will act as a barrier to
technology transfer.
 PHM-753
Syeda Gull-e Fatima
MPhil 1st
Applications Of Biotechnology Products
Along With Marketed Products
 Recombinant proteins

 Recombinant Erythropoietin Applications

Erythropoietin is an endogenous hormone produced in the kidney that regulates red blood cell
production within the body. Anemic people suffer from tissue hypoxia. Severe cases can
require transfusion of red cells from blood donors. Transfusion therapy with allogenic blood
components may cause immunologic reactions and infections. In addition, repeated red blood
cell transfusions can lead to iron overload. Therefore, the availability of recombinant human
erythropoietin (rHuEPO) as an antianemic drug has been an important medical progress.
 Available forms
 Epoetin alfa and epoetin beta, are currently used for treatment of EPOdeficiency anemia
and for support in autologous blood collection programs.
 Human Albumin 20%
by Biotest
 Human Albumin Biotest 20% is a solution for infusion (into a vein). 1 litre solution
contains 200 g human plasma protein of which at least 96% is human albumin.
 Human Albumin Biotest 20% is used to restore and maintain circulating blood volume
where there is a low
 blood volume and the use of a colloid, such as albumin, is required.
 https://www.medicines.org.uk/emc/files/pil.577.pdf
 Monoclonal Antibodies

Monoclonal antibodies are extensively used in clinically to the diagnosis and

therapy of an array of human disorders, including cancer and infectious
diseases, and have been used for the modulation of immune responses.
 Marketed Product
Actemra (80,200,400) concentrate solution for infusion by Roche that contains
tocilizumab.
 Hormones

 Insulin
 1 toujeo(insulin glargine) 300units/ml (SC)
3 solostar pens (soln for injection in prefilled pen) long acting insulin

https://www.diabetesresearchclinicalpractice.com/a
rticle/S0168-8227(16)30265-0/fulltext
 probiotics
 ’ The Food and Agriculture Organization of United Nations (FAO) and
World Health Organization (WHO) define probiotics used in food as
‘Live microorganisms which when administered in adequate amounts
confer a health benefit on the host’
 Application of beneficial bacteria to protect against detrimental
bacteria in the gastrointestinal tract and thus reap a positive health
benefit is the basis of probiotic therapy.
 Commercial products
 Most of the commercial probiotic products consist of combinations of
probiotic strains and are available in the form of powder, capsule, tablets,
drops, chewing gum, lozenges, straws, stick packs, bottle caps, etc. In
addition to probiotic strains, supplement formulations may also be added with
vitamins and prebiotics. Some probiotics meant for infants are sold in oil
suspensions making them easier to administer to infants. Also, use of
probiotics along with oral rehydration salts is an option for the treatment of
acute diarrhea.
 Marketed Examples
 Actiflor sachet by pharmevo( saccharomyces boulardii)
Treatment of infectious and non specific diarrhea. Also used in
combination with oral re hydration for acute diarrhea.
 Recombinant vacci nes
 Clinical Diagnostic Applications

 Clinical diagnostics applications include:

• Autoimmunity testing
• Blood typing and blood-borne disease assays
• Genetic testing
• Microbiology testing
• Recombinant protein purification
• IgG purification and virus removal
• Monoclonal antibody (mAb) purification
• Purification of serum proteins and enzymes
https://www.bio-rad.com/featured/en/biotechnology-equipment.html