Anticorpo monoclonal contra uma proteína macho-específica de 19 kDa em espermatozóides bovinos: uma metodologia promissora para imunosexagem

Abstract

RESUMO -Neste trabalho, foi purificada uma proteína macho específica com peso molecular de 19 Kda, a partir de espermatozóides de bovinos, por intermédio de cromatografia de gel filtração. O anticorpo monoclonal (Mab) desenvolvido (C11F) contra esta proteína discriminou duas populações de espermatozóides, como revelado por imunofluorescência em microscopia e citometria de fluxo. O epitopo reconhecido pelo Mab C11F esteve presente em 49,7% da população de espermatozóides. Quando adicionado ao sêmen bovino fresco, o Mab C11F reduziu a motilidade e promoveu a aglutinação dos espermatozóides.Palavras-chave: espermatozóides, bovinos, imunosexagem, proteína macho específica, ac. monoclonal Monoclonal Antibody against Male-Specific Protein of 19 KDa from BovineSpermatozoa: A Successful Methodology for Imunosexing ABSTRACT -In this work, a 19KDa, male specific protein, from bovine spermatozoa, was purified using gel filtration chromatography. A monoclonal antibody (Mab) developed (C11F) against this protein discriminated two populations of spermatozoa, as revealed by immunofluorescence in microscopy and flow citometry. The epitope recognized by Mab C11F was present in 49.7% of spermatozoa population. When added to fresh bovine semen, the Mab C11F reduced the motility and promoted agglutination of the spermatozoa.

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

“…Previous studies have employed hybridoma technology or various strategies of immunization aiming at producing antibodies against specific membrane proteins on sperm cells (Srinivasan et al, 1995;Souza et al, 1999;Blecher et al, 1999). Owing to their ability to specifically recognize antigens, monoclonal antibodies serve as important reagents in establishing protein identity (Liu et al, 2002).…”

“…Owing to their ability to specifically recognize antigens, monoclonal antibodies serve as important reagents in establishing protein identity (Liu et al, 2002). Antibodies have been used to identify sperm antigens binding to zona pellucida (Sivapurapu et al, 2003;Kaul et al, 2005), to isolate capacitation-related antigens (Mollova et al, 1999), and to identify sexspecific antigens (Souza et al, 1999;Blecher et al, 1999) and other antigens with interesting functions. Using an antibody phage display library, monoclonal antibodies against different sperm proteins were obtained in a less laborious and time-consuming manner.…”

“…On the other hand, Hendriksen et al (1996) cited some examples demonstrating that this exchange does not occur for all gene products. Souza et al (1999) purified and produced a monoclonal antibody against a 19-kDa bovine sperm membrane that bound to ∼50% of the sperm population by flow cytometer and only male tissues were staining by immunohistochemistry. Later, Matta et al (2001) obtained 92.3 and 80% of female embryos by ICSI and IVF, respectively, using monoclonal antibody produced by Souza et al (1999).…”

“…Souza et al (1999) purified and produced a monoclonal antibody against a 19-kDa bovine sperm membrane that bound to ∼50% of the sperm population by flow cytometer and only male tissues were staining by immunohistochemistry. Later, Matta et al (2001) obtained 92.3 and 80% of female embryos by ICSI and IVF, respectively, using monoclonal antibody produced by Souza et al (1999). Our results also suggest the presence of antigens characteristic of subpopulations of sperm cells and these antigens are apparently sex-specific.…”