Production of monoclonal antibodies against recombinant human zona pellucida glycoproteins: utility in immunolocalization of respective zona proteins in ovarian follicles

Bukovsky, Antonin; Gupta, Satish K.; Bansal, Pankaj; Chakravarty, Sanchita; Chaudhary, Manjari; Svetlikova, Marta; White, Rachel S.; Copas, Pleas; Upadhyaya, Nirmala B.; Van Meter, Stuart E.; Caudle, Michael R.

doi:10.1016/j.jri.2007.10.004

« Previous Next »Journal of Reproductive Immunology
Volume 78, Issue 2 , Pages 102-114, July 2008

Production of monoclonal antibodies against recombinant human zona pellucida glycoproteins: utility in immunolocalization of respective zona proteins in ovarian follicles

Antonin Bukovsky
- Laboratory of Development, Differentiation and Cancer, The University of Tennessee Graduate School of Medicine, 1924 Alcoa Highway, Knoxville, Tennessee 37920, USA
- Department of Obstetrics and Gynecology, The University of Tennessee Graduate School of Medicine, Tennessee, USA
- These authors contributed equally to this work.
- Corresponding author at: Laboratory of Development, Differentiation and Cancer, The University of Tennessee Graduate School of Medicine, 1924 Alcoa Highway, Knoxville, Tennessee 37920, USA. Tel.: +1 865 544 8969; fax: +1 865 544 6105.
Satish K. Gupta
- Gamete Antigen Laboratory, National Institute of Immunology, New Delhi, India
- These authors contributed equally to this work.
Pankaj Bansal
- Gamete Antigen Laboratory, National Institute of Immunology, New Delhi, India
Sanchita Chakravarty
- Gamete Antigen Laboratory, National Institute of Immunology, New Delhi, India
Manjari Chaudhary
- Gamete Antigen Laboratory, National Institute of Immunology, New Delhi, India
Marta Svetlikova
- Laboratory of Development, Differentiation and Cancer, The University of Tennessee Graduate School of Medicine, 1924 Alcoa Highway, Knoxville, Tennessee 37920, USA
- Department of Obstetrics and Gynecology, The University of Tennessee Graduate School of Medicine, Tennessee, USA
Rachel S. White
- Laboratory of Development, Differentiation and Cancer, The University of Tennessee Graduate School of Medicine, 1924 Alcoa Highway, Knoxville, Tennessee 37920, USA
- Department of Obstetrics and Gynecology, The University of Tennessee Graduate School of Medicine, Tennessee, USA
Pleas Copas
- Department of Obstetrics and Gynecology, The University of Tennessee Graduate School of Medicine, Tennessee, USA
Nirmala B. Upadhyaya
- Department of Obstetrics and Gynecology, The University of Tennessee Graduate School of Medicine, Tennessee, USA
Stuart E. Van Meter
- Department of Pathology, The University of Tennessee Graduate School of Medicine, Knoxville, Tennessee 37920, USA
Michael R. Caudle
- Laboratory of Development, Differentiation and Cancer, The University of Tennessee Graduate School of Medicine, 1924 Alcoa Highway, Knoxville, Tennessee 37920, USA

Received 26 April 2007; received in revised form 17 September 2007; accepted 22 October 2007. published online 04 March 2008.

Abstract

The zona pellucida (ZP) glycoproteins play an important role in oocyte development and gamete biology. To analyze their expression in follicles during various developmental stages, murine monoclonal antibodies (MAbs) were generated against the baculovirus-expressed recombinant human ZP2, ZP3 and ZP4. A panel of MAbs specific for the respective zona protein in ELISA and Western blot, and devoid of cross-reaction with other zona proteins was selected. Immunohistochemistry has shown that ZP2 MAb, MA-1620, did not react with oocytes in resting primordial follicles but showed reactivity with degenerating oocytes in primordial follicles undergoing atresia, and with oocytes in growing and antral follicles. Three MAbs against ZP3 did not react with oocytes in primordial follicles, but reacted only with oocytes in growing and antral follicles. Out of four MAbs against ZP4, three MAbs reacted with oocytes in primordial, growing and antral follicles. No reactivity of these MAbs with other ovarian cell types and other tissues studied (endometrium, uterine cervix, fallopian tubes and kidney) was detected except for a strong reactivity of ZP2 MA-1620 with epithelial cells of the uterine ectocervix or endometrium in some samples investigated. Altogether, these studies document generation of MAbs exhibiting high specificity for human zona proteins, which will be useful reagents to study their immunobiology.

Keywords: Zona pellucida, Oocyte development, Gamete biology, Monoclonal antibodies