Surface plasmon resonance detection of interactions between peptide fragments of N-telopeptide and its monoclonal antibodies.
As populations age, osteoporosis is becoming an important public health care problem. Urinary level of the cross-linked N-telopeptide of type I collagen has been reported to be a sensitive marker of bone resorption. Recently, we synthesized and characterized 10 overlapping peptides covering the N-telopeptide of alpha-2 type I collagen and reported their relative binding response to anti-type I collagen cross-linked N-telopeptide (NTX) antibodies determined by a competitive-inhibition enzyme-linked immunosorbent assay (ELISA). In this study, we design an assay based on the surface plasmon resonance (SPR) technology to detect binding interaction of each peptide fragment of NTX with the anti-NTX monoclonal antibodies. Anti-NTX monoclonal antibodies were immobilized on the surface of sensor chip by amine-coupling procedure. Serial dilutions of each peptide were prepared and injected separately onto the antibodies-immobilized sensor chip. The real-time association and dissociation interactions of each peptide were detected and reported as sensorgrams. Binding response of each peptide to the monoclonal antibodies was determined, and the SPR results were compared with the ELISA results. We demonstrate that the trends of binding potency of peptide fragments detected by SPR are in good correlation to the results obtained by ELISA, indicating that our developed SPR-based method can be further applied to detect the NTX fragments in urine and to monitor the bone loss in humans. The potent peptide fragments identified by both assays are promising for further preparation of specific monoclonal antibodies in order to develop bioassays for bone loss in humans.
Lung
F-D T
FD
Department of Nutrition, China Medical University, Taichung, Taiwan. fdlung@mail.cmc.edu.tw
Chen
C-H
CH
Liou
C-C
CC
Chen
H-Y
HY
D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D001666:Binding Sites, Antibody; D015415:Biomarkers / Q000652:urine; D003094:Collagen / Q000737:chemistry* / Q000276:immunology; D024042:Collagen Type I; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D010024:Osteoporosis / Q000175:diagnosis / Q000652:urine; D010446:Peptide Fragments / Q000737:chemistry* / Q000276:immunology / Q000652:urine; D011237:Predictive Value of Tests; D020349:Surface Plasmon Resonance
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D024042:Collagen Type I; D010446:Peptide Fragments; D003094:Collagen