Development of Immunoassay Using Monoclonal Antibody and Recombinant Antibody for Natural Products

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Hiroyuki Tanaka

Abstract

Introduction: Artemisinin (AM), which is a sesquiterpene lactone, is a well-known anti-mala rial compound isolated from Artemisia annua. We had prepared a monoclonal antibody (mAb 1C1) showing specifi city for AM and artesunate (AS), and we developed an indirect competitive enzymelinked immunosorbent assay (icELISA) using this novel mAb. Moreover, a recombinant antibody derived
from mAb 1C1 was prepared to overcome insuffi cient mAb production by hybridoma culture. Method: The recombinant antibodies, which were derived from a monoclonal antibody against AM and AS (mAb 1C1) prepared by us, were expressed by Escherichia coli cells, and their reactivity and specifi city were characterized. Results: As a result, the specifi city of the Fab was similar to that of mAb 1C1 in that it showed specifi c reactivity toward AM and AS only. The sensitivity of the icELISA (0.16 μg/mL – 40 μg/
mL for AM, 8.0 ng/mL – 60 ng/mL for AS) was suffi cient for analysis of anti-malarial drugs, and its utility for quality control of analysis of Artemisia spp. was validated. The Fab expression and refolding systems provided a good yield of high-quality antibodies.

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