• Journal of Sensor Science and Technology
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• v.18 no.2
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• pp.160-167
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• 2009

In this study, secondary antibody-containing quartz crystal microbalance(QCM) sensor chip was prepared and utilized for the detection of human and bovine haptoglobin. Anti-goat immunoglobulin G antibody, which is a secondary antibody capable of capturing primary antibodies raised in goat, was immobilized through the reaction between hydrazide and aldehyde group prepared on the QCM surface and antibody respectively. The resulting sensor chip showed higher stability in the repeated surface regeneration with acidic dissociation solution as well as requiring lower amount of primary antibody when compared to the protein G sensor chip. The secondary antibody sensor chip was applied for the estimation of bovine and human haptoglobin.

• BMB Reports
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• v.40 no.6
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• pp.1028-1038
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• 2007

Human plasma Hp is classified as 1-1, 2-1, and 2-2. They are inherited from two alleles Hp 1 and Hp 2, but there is only Hp 1 in almost all the animal species. Hp 2-2 molecule is extremely large and heterogeneous associated with the development of inflammatory-related diseases. In this study, we expressed entire bovine Hp in E. coli as a $\alpha\beta$ linear form. Interestingly, the antibodies prepared against this form could recognize the subunit of native Hp. In stead of a complicated column method, the antibody was able to isolate bovine Hp via immunoaffinity and gelfiltration columns. The isolated Hp is polymeric containing two major molecular forms (660 and 730 kDa). Their size and hemoglobin binding complex are significantly larger than that of human Hp 2-2. The amino-acid sequence deducted from the nucleotide sequence is similar to human Hp 2 containing a tandem repeat over the $\alpha$ chain. Thus, the Hp 2 allele is not unique in human. We also found that there is one additional -SH group (Cys-97) in bovine $\alpha$ chain with a total of 8 -SH groups, which may be responsible for the overall polymeric structure that is markedly different from human Hp 2-2. The significance of the finding and its relationship to structural evolution are also discussed.