title: Isolation of antibodies against different protein conformations using immunoaffinity chromatography.
abstract: A polyclonal antiserum obtained after the immunization of a rabbit with recombinant human sperm-specific glyceraldehyde-3-phosphate dehydrogenase lacking in 68 N-terminal amino acid residues (dN-GAPDS) was purified using different immunosorbents with immobilized dN-GAPDS in the native or denatured states. The procedure resulted in isolation of two types of polyclonal antibodies. The first type interacted with native recombinant dN-GAPDS as well as with native human sperm-specific glyceraldehyde-3-phosphate dehydrogenase, not cross-reacting with muscle glyceraldehyde-3-phosphate dehydrogenase (GAPD). The second type interacted with both native and denatured forms of the sperm-specific proteins, exhibiting some cross-reaction with GAPD. Thus, the suggested approach allows isolation of the antibodies against conformational or linear epitopes from the same polyclonal serum.
authors.affiliation: Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, 119991 Moscow, Russian Federation.
authors.forename: Mikhail L
authors.initials: ML
authors.lastname: Kuravsky
authors.forename: Elena V
authors.initials: EV
authors.lastname: Schmalhausen
authors.forename: Natalia V
authors.initials: NV
authors.lastname: Pozdnyakova
authors.forename: Vladimir I
authors.initials: VI
authors.lastname: Muronetz
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D000939:Epitopes / Q000276:immunology; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000276:immunology / Q000378:metabolism; D006801:Humans; D055767:Immobilized Proteins / Q000276:immunology / Q000378:metabolism; D007164:Immunosorbents / Q000276:immunology / Q000378:metabolism; D008297:Male; D009132:Muscles / Q000201:enzymology; D011489:Protein Denaturation; D011817:Rabbits; D013094:Spermatozoa / Q000201:enzymology
chemical_list: D000906:Antibodies; D000939:Epitopes; D055767:Immobilized Proteins; D007164:Immunosorbents; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases