Development and immunochemical evaluation of antibodies Y for the poorly immunogenic polypeptide prothymosin alpha.
Since conserved mammalian polypeptides are believed to exhibit enhanced immunogenicity in avian species, hens were immunized against the poorly immunogenic, highly conserved mammalian polypeptide prothymosin alpha (ProTalpha), i.e. against either non-conjugated ProTalpha (isolated from bovine thymus) or ProTalpha conjugated to keyhole limpet hemocyanin (ProTalpha/KLH). The antibodies Y were isolated from the egg yolk and evaluated through suitable dot-blot and ELISA systems in parallel with antibodies G isolated from the antiserum of rabbits immunized against the same immunogens. As revealed, antibodies Y and G of low titer and/or affinity were obtained against non-conjugated ProTalpha, while antibodies Y against ProTalpha/KLH had a better apparent titer, could better discriminate between ProTalpha and the closely related bioactive peptide thymosin alpha 1, and were obtained at much larger quantities than the corresponding antibodies G.
Klimentzou
Persefoni
P
National Centre for Scientific Research (NCSR) Demokritos, Institute of Radioisotopes & Radiodiagnostic Products, Immunopeptide Chemistry Laboratory, Aghia Paraskevi Attikis, Athens 15310, Greece.
Paravatou-Petsotas
Maria
M
Zikos
Christos
C
Beck
Alexander
A
Skopeliti
Margarita
M
Czarnecki
Jan
J
Tsitsilonis
Ourania
O
Voelter
Wolfgang
W
Livaniou
Evangelia
E
Evangelatos
Gregory P
GP
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002645:Chickens / Q000276:immunology*; D005260:Female; D006367:HeLa Cells; D006433:Hemocyanins / Q000276:immunology / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D007143:Immunoglobulin Heavy Chains / Q000302:isolation & purification; D007147:Immunoglobulin Light Chains / Q000302:isolation & purification; D007136:Immunoglobulins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology; D011498:Protein Precursors / Q000276:immunology* / Q000378:metabolism; D011817:Rabbits; D000077596:Thymalfasin; D013947:Thymosin / Q000031:analogs & derivatives* / Q000276:immunology / Q000378:metabolism
C016512:IgY; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007136:Immunoglobulins; D010446:Peptide Fragments; D011498:Protein Precursors; C040782:prothymosin alpha; D013947:Thymosin; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin; D000077596:Thymalfasin