DEVELOPMENT,PRODUCTION AND

OPTIMIZATION OF BLOOD GROUP

REAGENTS

SWETHA.R (2017401021)
M.TECH -BIOPHARMACEUTICAL
TECHNOLOGY
ANNA UNIVERSITY, CHENNAI.

External Guide
Internal Guide
Dr.Ganesh Prasad
Dr.Ramalingam
Senior Scientist
Assistant Professor
Bhat-Biotech India
Anna University
Pvt.Ltd
Chennai.
Bangalore
 INTRODUCTION
ABO BLOOD GROUPING
• Discovered by Karl Landsteiner
• Controlled by single ABO gene
• Gene – glycosyltransferase
• Modifies carbohydrate content of RBC
• Blood grouping based on presence of A,B antigen on the
surface of RBC
CLINICAL SIGNIFICANCE :
• Blood transfusion
• Haemolytic disease of new born
Test procedure for blood grouping reagents as per NIB,
• Titre
• Avidity
• Intensity
• Specificity
• Reactivity – Rouleux, prozone and hemolysis
 OBJECTIVES
• Development of monoclonal antibodies against blood group
antigens
• Characterization and quality tests of monoclonal antibodies
• Production Optimization and scale up
 WORK FLOW
Collection of blood samples for blood group antigens

Selection and immunization of mice

Fusion by hybridoma technology

Selection of clones by limited dilution method

Production of monoclonal antibodies against blood antigens

Characterization of antibodies

Production Optimisation and scale up

 WORK DONE

COLLECTION OF SAMPLES:
Samples for blood group antigens A,B and AB are collected from
different blood group individuals.
IMMUNIZATION:
20 % of blood 500μl for A antigen
No of mice: 6
IMMUNIZATION SCHEDULE:
Every alternative day for 24 days
HYBRIDOMA TECHNOLOGY
 RESULTS
SCREENING : BEFORE FUSION

Titer : 1:32
Titer: 1:64 Titer: 1:256
FUSION PLATES
SCREENING: AFTER FUSION

Titre: 1:64 Titre : 1:16

 RESULTS AND DISCUSSION
• The fusion plates did not show good results by
agglutination tests.
• Further optimization should be done for fusion by
hybridoma technology.
• After positive results are obtained for all the blood
grouping clones, characterization is done as per in
manual of National institute of biologics.
• Then production is optimized for scale up.
THANK YOU!!!!