Target antigens for Hs-14 monoclonal antibody and their various expression in normozoospermic and asthenozoospermic men

Basic and Clinical Andrology

Jana Capkova²  Hasmik Margaryan²  Alena Kubatova²  Petr Novak¹  Jana Peknicova² 

[1] Institute of Microbiology AS CR, v.v.i., Videnska 1083, Prague 4, 142 20, Czech Republic

[2] Laboratory of Reproductive Biology, Institute of Biotechnology AS CR, the Czech Academy of Sciences, v.v.i., Videnska 1083, Prague 4, 142 20, Czech Republic

关键词: Transitional endoplasmic reticulum ATPase;    Asthenozoospermia;    Monoclonal antibody;    Human spermatozoa;    Acrosome;   

Others  :  1230944
DOI  :  10.1186/s12610-015-0025-0

 received in 2015-06-10, accepted in 2015-10-22,  发布年份 2015

【 摘 要 】

Background

Poor semen quality is one of the main causes of infertility. We have generated a set of monoclonal antibodies to human sperm and used them to investigate sperm quality. Some of these antibodies found differences in the expression of proteins between normal sperm and pathological sperm displaying severe defects. One of them was the Hs-14 antibody.

The aim of this paper was to determine the target protein of the Hs-14 monoclonal antibody and to investigate the expression of the Hs-14-reacting protein on the sperm of asthenozoospermic men with sperm motility defect and of healthy normozoospermic men.

Methods

Indirect immunofluorescence, one-dimensional and two-dimensional polyacrylamide gel electrophoresis, immunoblotting and mass spectrometry.

Results

The Hs-14 antibody binds fibronectin, β-tubulin and valosin-containing protein - new name for this protein is transitional endoplasmic reticulum ATPase (TERA). Since the Hs-14 reaction with TERA remained the strongest at the highest antibody dilution, and Hs-14 consistently labelled the same spot or band as the monospecific anti-TERA antibody on immunoblots, we assume that TERA is an Hs-14-specific protein. Binding of fibronectin and β-tubulin might represent nonspecific cross-reactivity or Hs-14 reaction with similar epitopes of these proteins.

A significant difference (P < 0.001) in immunofluorescence staining with Hs-14 was found between the normozoospermic and asthenozoospermic men.

Conclusion

The Hs-14 antibody enables discrimination between sterile or subfertile asthenozoospermic and fertile normozoospermic men. Decreased levels of TERA in men can be used as a biomarker of reduced fertility.

【 授权许可】

   
2015 Capkova et al.

【 图 表 】

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