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Jung, Lee, Yong, and Cha: Isolation of Human scFv Antibodies Specific for House Dust Mite Antigens from an Asthma Patient by Using a Phage Display Library
Original Article

Immune Network 2002; 2(2): 91-95.

Published online: 30 June 2002

DOI: https://doi.org/10.4110/in.2002.2.2.91

Isolation of Human scFv Antibodies Specific for House Dust Mite Antigens from an Asthma Patient by Using a Phage Display Library

Wang lim Jung1, Hee kyung Lee1, Tae Soon Yong2, Sang Hoon Cha1, 3

1Division of Food Sci. & Biotechnology, College of Agriculture & Life Sci., Kangwon National University, Chunchon, Korea.

2Department of Parasitology, Yonsei University College of Medicine, Seoul, Korea.

3IG Therapy Co., Kangwon National University, Nong 3rd Rm. 112, Chuncheon, Korea.

Corresponding Author (chash@kangwon)

Copyright © 2002 The Korean Association of Immunologists

(open-access):

This is an open access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

BACKGROUND

In order to characterize human antibodies with specificity for mite allergens at the molecular level, a scFv phage display library was constructed using peripheral blood mononuclear lymphocytes from an asthma patient allergic to mite as Ig gene sources.

METHODS

Immunoglobulin VH and V gene fragments were obtained by polymerase chain reaction, and randomly combined in pCANTAB-5E vector. The resulting human scFv phage display library had 3 X 10(4) independent clones, and biopanning was performed with house dust mite extracts.

RESULTS

Four scFv clones specific for house dust mite extract were isolated. Immunoblot assay showed that our clones reacted to 25 kDa and 50~60 kDa proteins with unknown identity in mite extracts. Sequence analysis indicated that two clones (b7 and c15) are identical, and all clones belong to human VH3 subgroup. On the other hand, light chain usage was different in that two clones (a2 and b7/c15) belonging to V kappa 4 subgroup, but a4 used V kappa 1 light chain gene.

CONCLUSION

Our approach should facilitate provision of useful information on the antibody responses against allergens at the molecular level in humans.

Keywords: Phage display library, scFv, asthma, type-1 hypersensitivity, house dust mite, allergen

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