Production of monoclonal antibodies against recombinant human zona pellucida glycoproteins

utility in immunolocalization of respective zona proteins in ovarian follicles

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

Research output: Contribution to journalArticle

22 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)102-114
Number of pages13
JournalJournal of Reproductive Immunology
Volume78
Issue number2
DOIs
StatePublished - Jul 1 2008
Externally publishedYes

Fingerprint

Ovarian Follicle
Herpes Zoster
Oocytes
Monoclonal Antibodies
Proteins
Endometrium
Fallopian Tubes
Baculoviridae
Cross Reactions
Zona Pellucida Glycoproteins
Germ Cells
Cervix Uteri
Western Blotting
Epithelial Cells
Enzyme-Linked Immunosorbent Assay
Immunohistochemistry
Kidney
Antral

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology
  • Reproductive Medicine
  • Obstetrics and Gynecology

Cite this

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; Van Meter, Stuart; Caudle, Michael.

In: Journal of Reproductive Immunology, Vol. 78, No. 2, 01.07.2008, p. 102-114.

Research output: Contribution to journalArticle

Bukovsky, Antonin ; Gupta, Satish K. ; Bansal, Pankaj ; Chakravarty, Sanchita ; Chaudhary, Manjari ; Svetlikova, Marta ; White, Rachel S. ; Copas, Pleas ; Upadhyaya, Nirmala ; Van Meter, Stuart ; Caudle, Michael. / Production of monoclonal antibodies against recombinant human zona pellucida glycoproteins : utility in immunolocalization of respective zona proteins in ovarian follicles. In: Journal of Reproductive Immunology. 2008 ; Vol. 78, No. 2. pp. 102-114.
@article{4f2a94f5ff9f482db2fe5bff78407790,
title = "Production of monoclonal antibodies against recombinant human zona pellucida glycoproteins: utility in immunolocalization of respective zona proteins in ovarian follicles",
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.",
author = "Antonin Bukovsky and Gupta, {Satish K.} and Pankaj Bansal and Sanchita Chakravarty and Manjari Chaudhary and Marta Svetlikova and White, {Rachel S.} and Pleas Copas and Nirmala Upadhyaya and {Van Meter}, Stuart and Michael Caudle",
year = "2008",
month = "7",
day = "1",
doi = "10.1016/j.jri.2007.10.004",
language = "English (US)",
volume = "78",
pages = "102--114",
journal = "Journal of Reproductive Immunology",
issn = "0165-0378",
publisher = "Elsevier Ireland Ltd",
number = "2",

}

TY - JOUR

T1 - Production of monoclonal antibodies against recombinant human zona pellucida glycoproteins

T2 - utility in immunolocalization of respective zona proteins in ovarian follicles

AU - Bukovsky, Antonin

AU - Gupta, Satish K.

AU - Bansal, Pankaj

AU - Chakravarty, Sanchita

AU - Chaudhary, Manjari

AU - Svetlikova, Marta

AU - White, Rachel S.

AU - Copas, Pleas

AU - Upadhyaya, Nirmala

AU - Van Meter, Stuart

AU - Caudle, Michael

PY - 2008/7/1

Y1 - 2008/7/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=46549085543&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=46549085543&partnerID=8YFLogxK

U2 - 10.1016/j.jri.2007.10.004

DO - 10.1016/j.jri.2007.10.004

M3 - Article

VL - 78

SP - 102

EP - 114

JO - Journal of Reproductive Immunology

JF - Journal of Reproductive Immunology

SN - 0165-0378

IS - 2

ER -