Isolation and Characterization of Antichloramphenicol Antibodies using SDS Page

Jesca O Wesongah, Anastasia N Guantai

Abstract


Background: Antichloramphenicol antibodies can be produced in small or large animals depending on the requirement of the researcher. Previously most researchers have raised antibodies in small animals such as rabbits due to their easy availability and handling. In the present study antichloramphenicol antibodies were produced in large animals because large volumes of serum was needed for various studies.

Objective: The objective of the present study was to isolate and characterize antichloramphenicol antibodies produced in camels, donkeys and goats for development of a CAP Enzyme Linked Immunosorbent Assay.

Methods: The methods employed were SDS-PAGE electrophoresis which involved the analysis of crude and purified goat, camel and donkey antichloramphenicol antibodies. Purification of the antichloramphenicol antibodies was carried out by precipitation using ammonium sulphate. Immunization of experimental animals was carried out using standard immunological methods.

Results: The results indicated that the crude anti-CAP antibody produced in camels, goats and donkeys showed 7 protein bands of molecular sizes 11.7, 40, 61.6, 134.3, 145, 169.5 and 182 kda. However the protein band of molecular weight 11.7 kda was not observed in the purified antibody from the 3 animal species.  The protein bands of the camel appeared smaller and were more distinct as compared to those of donkeys and goats.

Conclusion: From this study it was concluded that purified camel antibodies are smaller and more specific followed closely by goat antibodies and donkey antibodies.

Keywords: anti-chloramphenicol (CAP) antibodies, camels, goats and donkeys


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