Surface plasmon resonance immuno assays - A perspective.
Human growth hormone (GH) represents an extremely challenging task from an anti-doping viewpoint. GH is an endogenously produced substance, present at very low levels in circulation (for the most abundant 22kDa isoform approximately 50pM in plasma and 100fM in urine) either as monomer or homo- and heterodimers, comprises a family of distinct isoforms, and obeys a pulsatile secretion routine that is affected by many different internal and external factors. Upon administration of the recombinant, single-isoform pharmaceutical, the feedback mechanism reduces the endogenous heterogeneity resulting in altered ratios between the different GH isoforms. Thus, measuring the isoform ratios through immuno assays appears the approach of choice. Conventional assays do not provide information on isoform-specific association and dissociation events of the individual primary antibody-isoform or isoform-secondary antibody interactions. This particular information can be obtained using the technology of surface plasmon resonance (SPR) which enables monitoring of biomolecular interactions in a dynamic and label-free setting. In this paper the different aspects of SPR are described, how the technology may be beneficial for understanding today's anti-GH immunoassays, and whether the approach could be employed for measuring GH in the near future.
Gutiérrez-Gallego
R
R
Bioanalysis and Analytical Services Research Group, Neuropsychopharmacology Program, Municipal Institute of Medical Research (IMIM-Hospital del Mar), PRBB, Barcelona, Spain.
Bosch
J
J
Such-Sanmartín
G
G
Segura
J
J
D000906:Antibodies / Q000737:chemistry; D015374:Biosensing Techniques; D019281:Dimerization; D004300:Doping in Sports; D019382:Human Growth Hormone / Q000032:analysis / Q000097:blood / Q000378:metabolism* / Q000652:urine; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007700:Kinetics; D008956:Models, Chemical; D010446:Peptide Fragments / Q000032:analysis / Q000097:blood* / Q000652:urine*; D040081:Protein Array Analysis; D011485:Protein Binding; D020033:Protein Isoforms; D015813:Substance Abuse Detection / Q000379:methods*; D020349:Surface Plasmon Resonance / Q000379:methods*
D000906:Antibodies; D010446:Peptide Fragments; D020033:Protein Isoforms; C404802:human growth hormone 22K; D019382:Human Growth Hormone