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Bioassay: Satish Kumar.J PH.D Student Mysore University

This document discusses bioassays, which involve using live organisms or tissues to determine the biological activity of a substance like a drug. It provides a brief history of bioassays dating back to the Bible and outlines their key purposes, principles, types, structure, advantages, and disadvantages. Bioassays can be used to standardize drugs and other materials when chemical methods are not available or effective. They allow determining potency and effects on organisms, but require many live subjects and expertise to perform.

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aziskf
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202 views

Bioassay: Satish Kumar.J PH.D Student Mysore University

This document discusses bioassays, which involve using live organisms or tissues to determine the biological activity of a substance like a drug. It provides a brief history of bioassays dating back to the Bible and outlines their key purposes, principles, types, structure, advantages, and disadvantages. Bioassays can be used to standardize drugs and other materials when chemical methods are not available or effective. They allow determining potency and effects on organisms, but require many live subjects and expertise to perform.

Uploaded by

aziskf
Copyright
© © All Rights Reserved
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
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Bioassay

SATISH KUMAR.J
PH.D STUDENT
MYSORE UNIVERSITY
Slide Title

Introduction
History of bioassay
Purpose of bioassay
Principle of bioassay
Structure of bioassay
Types of bioassay
Advantages of bioassay
Disadvantages of bioassay
Slide Title
• Bioassay (commonly used shorthand for biological
assay or assessment), or biological standardization
is a type of scientific experiment. A bioassay
involves the use of live animal or plant (in vivo) or
tissue or cell (in vitro) to determine the biological
activity of a substance, such as a hormone or drug
even a pollutants.
• Bioassays are typically conducted to measure the
effects of a substance on a living organism and are
essential in the development of new drugs and in
monitoring environmental pollutants.
• A bioassay can also be used to determine the
concentration of a particular constitution of a
mixture that may cause harmful effects on
organisms or the environment.
History of biological assay

In the Bible, in the description of Noah’s experiment


from his ark by sending a dove repeatedly until it
returns with an olive leaf, by which Noah knows or
estimates the level of receding waters from the Earth’s
grounds, we find that it has all the three essential
constituents of an assay – namely “stimulus” (depth of
water), “subject” (the done) and “response” (plucking of
an olive leaf).
Serious scientific history of biological assay began at the
close of 19th century with Ehrlich’s investigations into
the standardization of diphtheria antitoxin. Since then,
the standardization of materials by means of the
reactions of living matter has become a common
practice, not only in pharmacology, but in other
branches of science also, such as plant pathology and
environmental science.
However the assays were put on sound bases only since
1930’s when some statisticians contributed with their Father of chemotherapy
refined methods to this area.
Purpose of bioassay
Chemical method is either not available, if available
too complex or insensitive to low doses
If active principle of drug is not know
Unknown chemical composition
Active principle cannot be isolated
Not possible to separate inferring substance
Determination of the side-effect profile, including the
degree of drug toxicity.
Assessing the amount of pollutants being released by a
particular source, such as wastewater or urban runoff.
Principle of bioassay:

The bioassay compares the test sample with a same


Internationally applicable standard substance. It determines
the quantity of test sample required to produce an equivalent
biological response to that of standard substance.
Standard samples are accepted by expert committee at
international level and they represent fixed units of activity.
STRUCTURE OF BIOASSAY:

The typical bioassay involves a stimulus (for example, a vitamin, a drug, a


fungicide),applied to a subject(for example, an animal, a piece of animal tissue, a
plant, a bacterial culture).
The intensity of the stimulus is varied by using the various “doses” by the
experimenter.
Application of stimulus is followed by a change in some measurable characteristic of
the subject, the magnitude of the change being dependent upon the dose.
A measurement of this characteristic, for example, a weight of the whole subject, or of
some particular organ, an analytical value such as blood sugar content or bone ash
percentage, or even a simple record of occurrence or non-occurrence of a certain
muscular contraction, recovery from symptoms of a dietary deficiency, or death — is
the response of the subject
Types
Bioassays may be qualitative or quantitative.
Qualitative bioassays are used for assessing the physical effects of a substance that may
not be quantified, such as seeds fail to germinate or develop abnormally deformity.

An example of a qualitative bioassay includes Arnold Adolph Berthold's famous experiment


on castrated chickens. This analysis found that by removing the testicles of a chicken, it
would not develop into a rooster because the endocrine signals necessary for this process
were not available.

Quantitative bioassays involve estimation of the dose-response curve, how the response
changes with increasing dose. That dose-response relation allows estimation of the dose or
concentration of a substance associated with a specific biological response, such as the
LC50 (concentration killing 50% of the exposed organisms). Quantitative bioassays are
typically analyzed using the methods of biostatistics.
.
Graded Response Assay or quantitative bioassay.
Graded Response Assay: In these assays, as the dose increases there is an equivalent rise
in response. The potency is estimated by comparing the Test sample responses with the
standard response curve.
In the graded dose response relationship, relates the size of the response to the drug in a
single biologic unit as the dose administered increased the pharmacological response also
increases and eventually reaches a steady level called the ceiling effect there will be on
further increase in response even with an increase in dose.
The graded dose response curve is obtained by plotting a graph with dose on the X-axis
and response on the Y-axis. It is usually sigmoid in shape however the log dose response
curve is almost a straight line and particularly useful in bio assay.
Conc. of unknown= Threshold dose of standard/threshold dose of test x Conc. of
standard.
End Point or Quantal Assay: As the name indicates, the threshold dose of the
sample required to elicit a complete or a particular pharmacological effect is
determined and compared with standard.
E.g.,
TC (Tubocurarine) producing neck relaxation in rabbit, Here as the sample is
injected to the neck muscle of the Rabbit, the neck starts to droop. On further
doses there is complete hanging of the neck and rabbit has no ability to lift the
neck
Even the Determination of LD50 (LD=Lethal dose) or ED50 (ED= effective dose)
is done by this method.
Based on the method used during the grade point assay procedure for
determination of Type of activity and Potency of the Sample, four methods
of assays are classified as
a) Matching point or bracketing method
b) Interpolation assay
c) Three point (2+1) assay
d) Four- point (2+2) assay
Matching point or Bracketing method Here a constant dose of the standard is bracketed by
varying dose of sample until an exact matching between the standard dose responses and
the particular dose response of the sample is achieved.
To determine potency of test a log dose response curve is plotted.
Based on techniques they can be differentiated into two broad types like
a) In vivo techniques: These techniques employ a living animal recommended for the
purpose of assay. The techniques aims to study the biological effect or response of the
compound under screening in a living system directly. Ex: By use of rodents, rabbits etc.
b) In vitro techniques: These techniques employ a cell culture of recommended
biological system to study the effect of compound under standard condition not similar
to that of living environment. Here the cell culture survives by utilization of the
nutrition in the media. Ex: use of stem cells, cell culture, microbes (bacteria) etc
c) Ex vivo techniques: These techniques employ a tissue or cells of recommended living
system to study the effect of compound under test in suitable conditions within the
stipulated time of organ survival outside the body. The methods employ a living tissue
of an animal in an apparatus to study the contractile effect of drugs.
Ex: Use of any isolated organ from animals in a glass ware to study the effect of
compound within the period of its survival outside the living body with provision of only
oxygen, glucose and isotonic salts to maintain cell & cell organelles integrity.
Advantages & Uses of bioassay
1. They not only help to determine the concentration but also the potency of the
sample. (Potency is a term which denotes activity of the compound per molecule
basis. i.e. if a compound shows better activity at minute concentration, greater is
the potency, and if its activity is low at lower concentrations, lesser is the potency).
2.It is especially used to standardize drugs, vaccines, toxins or poisons,
disinfectants, antiseptics etc. as these are all used over biological system in some
or other form.
3. These also help determine the specificity of a compound to be used ex:
Penicillin's are effective against Gram+ve but not on Gram-ve. Testing of infected
patients sputum helps determine which anti-biotic be given for quick recovery.
4. Certain complex compounds like Vitamin B-12 which can't be analyzed by
simple assay techniques can be effectively estimated by Bioassays.
5. Sometimes the chemical composition of samples are different but have same
biological activity.
5. For samples where no other methods of assays are available
DISADVANTAGES OF BIOASSAY

Key problem is variable in response


Large number of animals to be used
Expertise in experimental design , execution of assay and analysis
of data required
leads to expensive and time consuming
Time related changes in sensitivity of test organ
REFERENCES

•www.math.iitb.ac.in/~ashish/workshop/gmsa
haw3.pdf
•https://en.wikipedia.org/wiki/Bioassay
studentstudyhub.com/wp-content/uploads/2013/11/Bioassay.docx
www.ncbi.nlm.nih.gov/pubmed/17837502

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