Analytical identification of additional impurities in urinary-derived gonadotrophins.
Advances in proteomic technology have enabled contaminant proteins to be identified from complex protein mixtures. The purity of two purified urinary gonadotrophin products, human menopausal gonadotrophin (u-HMG) and human FSH (u-hFSH), was compared with a preparation of recombinant human FSH (r-hFSH). After separation by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), western blot analysis showed that the recombinant preparation contained only FSH, whereas the urine-derived preparations exhibited several non-FSH or LH-related bands. These urinary components were further investigated by a proteomic approach using two-dimensional SDS-PAGE followed by mass spectrometric identification. The proteomic approach detected a total of 23 non-gonadotrophin-related proteins, at variable levels in different batches of the urine-derived preparations. Of these, 16 co-purified proteins have not been previously reported to be present in urine-derived gonadotrophins. These results indicate that the process used to purify urinary gonadotrophins may not remove all non-gonadotrophin proteins. By using a comprehensive proteomic approach, it has been shown that the recombinant FSH preparation has greater purity than either of the urine-derived preparations.
Bassett
R
R
Merck Serono S.A. - Geneva, 11 Chemin des Mines, Geneva 1201, Switzerland. robert.bassett@merckserono.net
Lispi
M
M
Ceccarelli
D
D
Grimaldi
L
L
Mancinelli
M
M
Martelli
F
F
Van Dorsselaer
A
A
D004340:Drug Contamination; D004341:Drug Evaluation; D004591:Electrophoresis, Polyacrylamide Gel; D005640:Follicle Stimulating Hormone / Q000032:analysis / Q000592:standards; D006801:Humans; D008596:Menotropins / Q000032:analysis* / Q000378:metabolism / Q000592:standards / Q000652:urine; D011506:Proteins / Q000302:isolation & purification / Q000378:metabolism; D040901:Proteomics; D011994:Recombinant Proteins / Q000032:analysis / Q000592:standards; D014556:Urine / Q000737:chemistry
D011506:Proteins; D011994:Recombinant Proteins; D008596:Menotropins; D005640:Follicle Stimulating Hormone