Application of recombinant antibody library for screening specific antigens in a bovine sperm cell subpopulation

• Published in: Livestock science Vol. 114; no. 2; pp. 188 - 193
• Main Authors: Soares, Sandro Gomes, Barbosa, Jose Elpidio
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01-04-2008; Amsterdam; New York: Elsevier
• Subjects: antibody detection; antibody phage display libraries; antigens; biomarkers; cattle; females; flow cytometry; leukocytes; males; membrane proteins; Nellore; Nelore; Phage display; proteomics; recombinant antibodies; recombinant antibody libraries; scFv; Sexing; Simmental; spermatozoa; Nelore; Phage display; scFv; Sexing
• ISSN: 1871-1413; 1878-0490
• DOI: 10.1016/j.livsci.2007.04.024

Antibody phage display libraries are a useful tool in proteomic analyses. This study evaluated an antibody recombinant library for identification of sex-specific proteins on the sperm cell surface. The Griffin.1 library was used to produce phage antibodies capable of recognizing membrane proteins from Nelore sperm cells. After producing soluble monoclonal scFv, clones were screened on Simental sperm cells by flow cytometry and those that bound to 40–60% of cells were selected. These clones were re-analyzed using Nelore sperm cells and all clones bound to 40–60% of cells. Positive clones were submitted to a binding assay against male and female bovine leukocytes by flow cytometry and one clone preferentially bound to male cells. The results indicate that phage display antibodies are an alternative method for identification of molecules markers on sperm cells.