Isolation and Characterization of Antichloramphenicol Antibodies using SDS Page

Jesca O Wesongah, Anastasia N Guantai


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


Daley LP, Gagliardo LF, Duffy MS, Smith MC and Appleton (2005). Application of monoclonal Antibodies in functional and Comparative Investigations of Heavy –Chain Immunoglobulins in New World Camelids. Clin, Diagn, Lab, Immunol. 12: 380-386.

Herrera M, Leon G; Segura A, Meneses F, Lomonte B, Chippaux JP and Gutierrez JM (2005) Factors associated with adverse reactions induced by caprylic acid fractionated whole IgG preparations comparison between horse, sheep and camel IgGs. Toxicon, 46:775-781.

Ladenson RC, Crmmins DL, Landt Y, and Landeson JH (2006) Isolation and characterization of a Thermally Stable Recombinant Anti-Caffeine Heavy–Chain Antibody Fragment. Anal. Chem. 78, 4501-4508.

Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680-685.

Lancaster M (2006). Veterinary drug residues and MRLS, 2006 Edition, ISBN: SR251.

Mohanty JG and Elazhary Y. (1989) Purification of IgG from serum with caprylic acid and ammonium sulphate precipitation is not superior to ammonium sulphate precipitation alone. Comp Immunol. Microb. Infect. Dis. 12:153-60.

Murilla GA (1996). Studies on trypanocidal drug, homidium, development and use of ELISA for its detection and quantification in cattle. PhD Thesis. University of Glasgow, United Kingdom.

Omidfar KR, Modjitahedi MJ, Forouzandeh H, Taghikhan M, Bakhtiari A, Paknejad M, and Kashanian S. (2004). Production and characterization of a new Antibody Specific for Mutant EGR receptor, EGFRvIII, in Camelus bactrianus. Tumor Biol. 25:179-187.

Wand M, Tran JH, Jacoby GA, Zhang Y, Wang F, and Hooper DC (2003). Plasmid-mediated quinolone resistance in clinical isolates of Escherichia coli from Shanghai China. Antimicrob Agents Chemother. 47: 2242-2248.

Wesongah JO, Guantai AN (2012) Potential Animal Sources of Antibodies for the development of a Chloramphenicol Enzyme-Linked Immunosorbent Assay. Afr. J. Pharmacol. Ther. 1:41-45.

Wesongah JO, Murilla GA, Guantai AN, Elliot C, Foddey T, and Cannavan A (2007). A competitive enzyme-linked immunosorbent assay for determination of chloramphenicol. J. of Vet. Pharmacol. Ther. 30, 1-6.

Zubritsky E (2005). Camel antibodies for PSA assys. Anal. Chem. 451 A:233-236.

Full Text: PDF


  • There are currently no refbacks.