Purification of human chorionic gonadotropin from pregnancy urine by immunoaffinity chromatography using a monoclonal antibody anti-beta chain hCG.
Human chorionic gonadotropin (hCG) is found in pregnancy urine and in urine from some cancer patients. It is found in a variety of forms, whose concentrations have clinical importance. A highly active preparation of hCG from pregnancy urine has been obtained by a simple method of immunoaffinity chromatography using a monoclonal antibody (MAb) against beta-subunit hCG. The recuperation of hCG from urine extract by this procedure was 84.18%, with a purification factor of 19.78. The specificity activity of the obtained hCG was 9705.36 IU/mg. The preparation appeared homogeneous by SDS-PAGE, analytical gel filtration, and double immunodiffusion. The molecular mass was 40 kDa by gel filtration on Superose 12 HR. Molecular masses estimated by electrophoresis were 14.6 and 23.5 kDa for the alpha and beta subunits, respectively.
Rodríguez
Antonio Melchor
AM
Laboratory of Protein Purification, Immunoassay Center, Cubanacán, Playa, Havana, Cuba. iqpurifica@ci,sld.cu
Rodríguez
Orlando Zulueta
OZ
Conde
Ileana Baluja
IB
Del Pino
Yanelis Robainas
YR
Pérez
Liliana Hernández
LH
Cisnero
Raquel López
RL
Duque
Maritza Gutierrez
MG
Mora
Julliette Cazanave
JC
Ariza
Dunia Bequer
DB
D000911:Antibodies, Monoclonal / Q000737:chemistry*; D006063:Chorionic Gonadotropin / Q000302:isolation & purification* / Q000652:urine; D018997:Chorionic Gonadotropin, beta Subunit, Human / Q000276:immunology; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D005260:Female; D006801:Humans; D011247:Pregnancy
D000911:Antibodies, Monoclonal; D006063:Chorionic Gonadotropin; D018997:Chorionic Gonadotropin, beta Subunit, Human