Construction of Antibody Library and Production of Polyclonal Antibodies Specific to Human Protein C
6
Issued Date
2023-06-01
Resource Type
ISSN
01252208
Scopus ID
2-s2.0-85164267785
Journal Title
Journal of the Medical Association of Thailand
Volume
106
Issue
6
Start Page
612
End Page
619
Rights Holder(s)
SCOPUS
Bibliographic Citation
Journal of the Medical Association of Thailand Vol.106 No.6 (2023) , 612-619
Suggested Citation
Prachasuphap A., Dhepakson P., Treeyoung P., Luengchaichawang A., Soonthorncharttrawat S., Jongpitisub K., Saengtong N., Kadegasem P., Sirachainan N. Construction of Antibody Library and Production of Polyclonal Antibodies Specific to Human Protein C. Journal of the Medical Association of Thailand Vol.106 No.6 (2023) , 612-619. 619. doi:10.35755/jmedassocthai.2023.06.13789 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/87966
Title
Construction of Antibody Library and Production of Polyclonal Antibodies Specific to Human Protein C
Author's Affiliation
Other Contributor(s)
Abstract
Background: Thromboembolism can occur at any age, from infancy to adulthood. The genetic risk factors for thromboembolism in the Thai population are protein C and protein S mutations. The diagnosis depends on the activity level of proteins in the plasma. Management of acute thrombotic events in severe protein C deficiency typically requires replacement with protein C in the form of fresh frozen plasma or protein C concentrate. At present, the measurement of protein C and protein S activities level is limited due to the cost and availability of the tests. Therefore, it can take a lengthy time to determine protein C levels, resulting in a delay in treatment, especially in severe cases. Objective: To create an antibody library on filamentous bacteriophage for generating antigen specific monoclonal antibodies and to produce polyclonal antibodies specific to protein C. Results: Recombinant protein C was constructed and produced from human embryonic kidney 293 cells. Protein C was used as an antigen to immunize mice and rabbits. The antibody titers after final boosting immunization reached up to 50,000 in mice and 10,000 in rabbits. The phage antibody library with a capacity of approximately 3×105 CFU was obtained from mice lymphocytes. Polyclonal antibodies were purified from rabbit sera using affinity chromatography and achieved a yield of 2 mg/mL rabbit sera. The purified polyclonal antibodies were able to detect protein C by immunoblotting and ELISA. Conclusion: The present study demonstrated that polyclonal antibodies could be produced from immunized rabbits and the antibody library obtained from immunized mice is beneficial for further isolation of monoclonal antibodies against protein C.