Characterization of monoclonal antibodies to glycodelin and recombinant glycodelin.
Glycodelin-A belongs to the lipocalin superfamily. Although it is associated with normal endometrial growth during the menstrual cycle, fertilization and normal pregnancy in humans, the molecular mechanism of its biological action has not been elucidated. To undertake studies to understand the functional relevance of any molecule, obtaining large quantities of the protein becomes essential. With the ultimate aim of purifying glycodelin either from its natural sources (human amniotic fluid) or the recombinant glycodelin from bacterial recombinant lysates, we raised monoclonal antibodies to this protein. As immunogens, recombinant glycodelin expressed in E. coli and Pichia pastoris as well as glycodelin from amniotic fluid were used. The monoclonal antibodies generated were characterized with respect to binding to both the native as well as the recombinant proteins using ELISA, immunoblotting, and immunohistochemistry.
Karri
S
S
Department of Biochemistry, Indian Institute of Science, Bangalore.
Mukhopadhyay
D
D
Jing
L
L
Karande
A A
AA
D000653:Amniotic Fluid / Q000737:chemistry; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000302:isolation & purification; D000915:Antibody Affinity; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D000071816:Glycodelin; D006023:Glycoproteins / Q000276:immunology*; D006801:Humans; D007132:Immunoglobulin Isotypes; D007150:Immunohistochemistry; D007166:Immunosuppressive Agents / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D010843:Pichia / Q000235:genetics; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000276:immunology*; D011994:Recombinant Proteins / Q000737:chemistry
D000911:Antibodies, Monoclonal; D000071816:Glycodelin; D006023:Glycoproteins; D007132:Immunoglobulin Isotypes; D007166:Immunosuppressive Agents; C045246:PAEP protein, human; C489047:Paep protein, mouse; D011257:Pregnancy Proteins; D011994:Recombinant Proteins