کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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1176618 | 961865 | 2012 | 8 صفحه PDF | دانلود رایگان |

Acute myocardial infarction is a typical disorder that requires continuous monitoring for early detection of potential life-threatening situations. To this end, we used different methods to screen for rapidly reversible antibodies, among 22 hybridoma clones, against cardiac troponin I (cTnI), which is a specific marker indicating the disease. The dissociation rates of antibodies were underestimated by up to a factor of 1000 because of bivalent binding when tested with the antigen immobilized on solid surfaces. This effect was also observed in a sandwich immunoassay, in which the detection antibody cross-linked with various antigen molecules already bound to the capture antibody. Although multiple binding events contributed to enhanced detection capability, it was difficult to recycle the immunosensor. We then devised a screening system by arranging the test antibody for the capture binder immobilized on a label-free sensor. This enabled us to select fast reactive antibodies of which one (clone 24) was shown to be recyclable, even in serum-containing medium. Using this antibody, repetitive detection of cTnI with a rapid response time (half-life of dissociation: about 4 min on average) and high detection capability (0.1 ng/ml) was achieved, which is very important for detection in a clinical setting.
Journal: Analytical Biochemistry - Volume 431, Issue 1, 1 December 2012, Pages 11–18