SDS-PAGE electrophoretic property of human chorionic gonadotropin (hCG) and its beta-subunit.
The microheterogeneity property of hCG with regards to its sialic acid contents resulted in variable mobility of the glycoprotein in SDS-PAGE. The intact hCG molecule is composed of two dissimilar subunits, namely alpha- and beta-subunits. The identification of hCG bands in SDS-PAGE was accomplished by the immunoblotting experiment, whereby the antibody directed toward the specific region of beta-subunit of hCG was used. The data shows that the different mobility of intact hCG was attributed to the different degree of desialylation of the glycoprotein. Nevertheless, unlike the intact hCG, the mobility of its beta-subunit was not affected by its variety sialic acid content. This characteristic of beta-hCG is beneficial when semi-quantification of total hCG is required. Quantification of hCG using the HPLC-reversed phase C18 analytical column is not possible as the glycoprotein was eluted in multiple fractions at different retention times. The identification of denatured hCG (HPLC eluted fractions) was carried out by immunoblotting experiment whilst immunoassay technique failed to detect its presence in any fraction.
Gam
Lay-Harn
LH
School of Pharmaceutical Sciences, University Sciences of Malaysia, 11800 Penang, Malaysia. layharn@usm.my
Latiff
Aishah
A
D015153:Blotting, Western; D006063:Chorionic Gonadotropin / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans
D006063:Chorionic Gonadotropin