• BMB Reports
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• v.42 no.11
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• pp.731-736
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• 2009

The generation of functional recombinant antibodies from hybridomas is necessary for antibody engineering. However, this is not easily accomplished due to high levels of aberrant heavy and light chain mRNAs, which require a highly selective technology that has proven complicated and difficult to operate. Herein, we attempt to use an alkaline phosphate (AP)-fused form of single-chain variable fragment (scFv) for the simple identification of a hybridoma-derived, functional recombinant antibody. As a representative example, we cloned the scFv gene from a hybridoma-producing mouse IgG against branched-chain keto acid dehydrogenase complex-E2 (BCKD-E2) into an expression vector containing an in-frame phoA gene. Functional recombinant antibodies were easily identified by conventional enzyme-linked immunosorbent assay (ELISA) by employing scFv-AP fusion protein, which also readily serves as a valuable immuno-detective reagent.

• Journal of The Korean Society of Inherited Metabolic disease
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• v.3 no.1
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• pp.4-14
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• 2003

Maple syrup urine disease or branched chain ketoacidurias caused by a deficiency in activity of the branched-chain ${\alpha}$-keto acid dehydrogenase(BCKD) complex. This metabolic block results in the accumulation of the branched-chain amino acids(BCAAs) leucine, isoleucine and valine, and the corresponding branched chain ${\alpha}$-keto acids (BCKAs). Based on the clinical presentation and biochemical responses to thiamine administration, MSUD patients can be divided into five phenotypes : classic, intermediate, intermittent, thiamine responsive and dihydrolipoyl dehydrogenase(E3)-deficient. Classic MSUD has a neonatal onset of encephalopathy, and is the most severe ad most common form. Variant forms of MSUD generally have the initial symptoms by 2 years of age. The majority of untreated classic patients die within the early months of life from recurrent metabolic crisis and neurologic deterioration. Treatment involves both longterm dietary management and aggressive intervention during acute metabolic decompensation. We report here our experience of longterm diet therapy and treatment of acute metabolic decompensation of a case of classic MSUD.