Phage display used for gene cloning of human recombinant antibody against the erythrocyte surface antigen, rhesus D

M Dziegiel; L K Nielsen; P S Andersen; A Blancher; E Dickmeiss; J Engberg

doi:10.1016/0022-1759(95)00013-Z

Phage display used for gene cloning of human recombinant antibody against the erythrocyte surface antigen, rhesus D

M Dziegiel, L K Nielsen, P S Andersen, A Blancher, E Dickmeiss, J Engberg

Publikation: Bidrag til tidsskrift › Tidsskriftsartikel › Forskning › peer review

Abstract

A novel phage display system has been developed for PCR amplification and cloning of the Fab fragments of human immunoglobulin genes. Using this system, we have cloned an antibody from a mouse-human hybridoma cell line directed against the erythrocyte antigen rhesus D. Intact erythrocytes were used for absorption of the Fab phages. Soluble Fab fragments produced from the cloned material showed identical performance to the parental antibody in agglutination assays. Gel filtration confirmed that the Fab fragment consists of a κ-Fd heterodimer. The successful use of intact cells for selection of specific Fab phages demonstrates that it is possible to by-pass purification of the antigen of interest. Comparison with published germline sequences demonstrated that the immunoglobulin coding regions had the highest homology to the V _H 1.9III and V _κ Humκv325 germline genes, respectively.

Originalsprog	Engelsk
Tidsskrift	Journal of Immunological Methods
Vol/bind	182
Udgave nummer	1
Sider (fra-til)	7-19
Antal sider	13
ISSN	0022-1759
DOI	https://doi.org/10.1016/0022-1759(95)00013-Z
Status	Udgivet - 11 maj 1995
Udgivet eksternt	Ja

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10.1016/0022-1759(95)00013-Z

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title = "Phage display used for gene cloning of human recombinant antibody against the erythrocyte surface antigen, rhesus D",

abstract = "A novel phage display system has been developed for PCR amplification and cloning of the Fab fragments of human immunoglobulin genes. Using this system, we have cloned an antibody from a mouse-human hybridoma cell line directed against the erythrocyte antigen rhesus D. Intact erythrocytes were used for absorption of the Fab phages. Soluble Fab fragments produced from the cloned material showed identical performance to the parental antibody in agglutination assays. Gel filtration confirmed that the Fab fragment consists of a κ-Fd heterodimer. The successful use of intact cells for selection of specific Fab phages demonstrates that it is possible to by-pass purification of the antigen of interest. Comparison with published germline sequences demonstrated that the immunoglobulin coding regions had the highest homology to the V H 1.9III and V κ Humκv325 germline genes, respectively.",

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author = "M Dziegiel and Nielsen, {L K} and Andersen, {P S} and A Blancher and E Dickmeiss and J Engberg",

year = "1995",

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AU - Dziegiel, M

AU - Nielsen, L K

AU - Andersen, P S

AU - Blancher, A

AU - Dickmeiss, E

AU - Engberg, J

PY - 1995/5/11

Y1 - 1995/5/11

N2 - A novel phage display system has been developed for PCR amplification and cloning of the Fab fragments of human immunoglobulin genes. Using this system, we have cloned an antibody from a mouse-human hybridoma cell line directed against the erythrocyte antigen rhesus D. Intact erythrocytes were used for absorption of the Fab phages. Soluble Fab fragments produced from the cloned material showed identical performance to the parental antibody in agglutination assays. Gel filtration confirmed that the Fab fragment consists of a κ-Fd heterodimer. The successful use of intact cells for selection of specific Fab phages demonstrates that it is possible to by-pass purification of the antigen of interest. Comparison with published germline sequences demonstrated that the immunoglobulin coding regions had the highest homology to the V H 1.9III and V κ Humκv325 germline genes, respectively.

AB - A novel phage display system has been developed for PCR amplification and cloning of the Fab fragments of human immunoglobulin genes. Using this system, we have cloned an antibody from a mouse-human hybridoma cell line directed against the erythrocyte antigen rhesus D. Intact erythrocytes were used for absorption of the Fab phages. Soluble Fab fragments produced from the cloned material showed identical performance to the parental antibody in agglutination assays. Gel filtration confirmed that the Fab fragment consists of a κ-Fd heterodimer. The successful use of intact cells for selection of specific Fab phages demonstrates that it is possible to by-pass purification of the antigen of interest. Comparison with published germline sequences demonstrated that the immunoglobulin coding regions had the highest homology to the V H 1.9III and V κ Humκv325 germline genes, respectively.

KW - Amino Acid Sequence

KW - Bacteriophages

KW - Base Sequence

KW - Cloning, Molecular

KW - DNA, Complementary

KW - Genetic Vectors

KW - Humans

KW - Isoantibodies

KW - Molecular Sequence Data

KW - Polymerase Chain Reaction

KW - Recombinant Fusion Proteins

KW - Rh-Hr Blood-Group System

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

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U2 - 10.1016/0022-1759(95)00013-Z

DO - 10.1016/0022-1759(95)00013-Z

M3 - Journal article

C2 - 7769246

SN - 0022-1759

VL - 182

SP - 7

EP - 19

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

IS - 1

ER -

Phage display used for gene cloning of human recombinant antibody against the erythrocyte surface antigen, rhesus D

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