High-resolution epitope mapping of hen egg lysozyme mabs by alanine-scanning mutagenesis

Yili Li, Jean-Charles B. Grivel, Malini Viswanathan, Smith-Gill

Research output: Contribution to journalArticle

Abstract

Protein-protein interactions are often modelled by antibody-antigen interactions and the understanding of molecular interactions underling antibody specificity and affinity are still to be resolved. \Ve have developed a mode! system of high affinity monoclonal antibodies specific for Hen Egg Lyso/yme (HEL). The structure of one of our antibodies HyHEL 10 complexed with H E I. has been defined by X-ray crystallography, and provide a definition of the st uctural epitope recognized by this antibody. The definition and the understanding of the interaction of this antibody with HEL require complementary sei of data, such as the definition of the residues contribul ing the energy of t ho interaction and defining the energetic epitope. Therefore the use of natural mutants or site directed mutants of both the antigen and the antibody is the too! of choice to answer these questions. HEL was actually used as prototypic antigen because of the numerous natural variants available, and we have designed and expressed around .'iO mutants of HEL in the methylatropic yeast pichia pastoris. 'I hese mutants obtained bv replacing residues near or at the interface of HyHEL 1ÜTIEI. complex with Ala, will be used to study the interaction with HyHKL 10 and also with two related antibodies of close specificity. HyHKL H and HyHEL 2fi. The sensitivity <>f these related antibodies to the mutants will also shed light on the contribution of antibody residues in the interaction, by correlating the sensitivities to the primary sequence of these diidtibodies.

Original languageEnglish
JournalFASEB Journal
Volume11
Issue number9
Publication statusPublished - 1997
Externally publishedYes

Fingerprint

Epitope Mapping
Mutagenesis
Alanine
Epitopes
Scanning
Antibodies
Ovum
Antibody Specificity
Antigens
Antibody Affinity
Pichia
X Ray Crystallography
hen egg lysozyme
Molecular interactions
X ray crystallography
Proteins
Yeasts
Monoclonal Antibodies
Yeast

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

High-resolution epitope mapping of hen egg lysozyme mabs by alanine-scanning mutagenesis. / Li, Yili; Grivel, Jean-Charles B.; Viswanathan, Malini; Smith-Gill.

In: FASEB Journal, Vol. 11, No. 9, 1997.

Research output: Contribution to journalArticle

@article{5ed6e153617e458c9f50764f447ed41f,
title = "High-resolution epitope mapping of hen egg lysozyme mabs by alanine-scanning mutagenesis",
abstract = "Protein-protein interactions are often modelled by antibody-antigen interactions and the understanding of molecular interactions underling antibody specificity and affinity are still to be resolved. \Ve have developed a mode! system of high affinity monoclonal antibodies specific for Hen Egg Lyso/yme (HEL). The structure of one of our antibodies HyHEL 10 complexed with H E I. has been defined by X-ray crystallography, and provide a definition of the st uctural epitope recognized by this antibody. The definition and the understanding of the interaction of this antibody with HEL require complementary sei of data, such as the definition of the residues contribul ing the energy of t ho interaction and defining the energetic epitope. Therefore the use of natural mutants or site directed mutants of both the antigen and the antibody is the too! of choice to answer these questions. HEL was actually used as prototypic antigen because of the numerous natural variants available, and we have designed and expressed around .'iO mutants of HEL in the methylatropic yeast pichia pastoris. 'I hese mutants obtained bv replacing residues near or at the interface of HyHEL 1{\"U}TIEI. complex with Ala, will be used to study the interaction with HyHKL 10 and also with two related antibodies of close specificity. HyHKL H and HyHEL 2fi. The sensitivity <>f these related antibodies to the mutants will also shed light on the contribution of antibody residues in the interaction, by correlating the sensitivities to the primary sequence of these diidtibodies.",
author = "Yili Li and Grivel, {Jean-Charles B.} and Malini Viswanathan and Smith-Gill",
year = "1997",
language = "English",
volume = "11",
journal = "FASEB Journal",
issn = "0892-6638",
publisher = "FASEB",
number = "9",

}

TY - JOUR

T1 - High-resolution epitope mapping of hen egg lysozyme mabs by alanine-scanning mutagenesis

AU - Li, Yili

AU - Grivel, Jean-Charles B.

AU - Viswanathan, Malini

AU - Smith-Gill,

PY - 1997

Y1 - 1997

N2 - Protein-protein interactions are often modelled by antibody-antigen interactions and the understanding of molecular interactions underling antibody specificity and affinity are still to be resolved. \Ve have developed a mode! system of high affinity monoclonal antibodies specific for Hen Egg Lyso/yme (HEL). The structure of one of our antibodies HyHEL 10 complexed with H E I. has been defined by X-ray crystallography, and provide a definition of the st uctural epitope recognized by this antibody. The definition and the understanding of the interaction of this antibody with HEL require complementary sei of data, such as the definition of the residues contribul ing the energy of t ho interaction and defining the energetic epitope. Therefore the use of natural mutants or site directed mutants of both the antigen and the antibody is the too! of choice to answer these questions. HEL was actually used as prototypic antigen because of the numerous natural variants available, and we have designed and expressed around .'iO mutants of HEL in the methylatropic yeast pichia pastoris. 'I hese mutants obtained bv replacing residues near or at the interface of HyHEL 1ÜTIEI. complex with Ala, will be used to study the interaction with HyHKL 10 and also with two related antibodies of close specificity. HyHKL H and HyHEL 2fi. The sensitivity <>f these related antibodies to the mutants will also shed light on the contribution of antibody residues in the interaction, by correlating the sensitivities to the primary sequence of these diidtibodies.

AB - Protein-protein interactions are often modelled by antibody-antigen interactions and the understanding of molecular interactions underling antibody specificity and affinity are still to be resolved. \Ve have developed a mode! system of high affinity monoclonal antibodies specific for Hen Egg Lyso/yme (HEL). The structure of one of our antibodies HyHEL 10 complexed with H E I. has been defined by X-ray crystallography, and provide a definition of the st uctural epitope recognized by this antibody. The definition and the understanding of the interaction of this antibody with HEL require complementary sei of data, such as the definition of the residues contribul ing the energy of t ho interaction and defining the energetic epitope. Therefore the use of natural mutants or site directed mutants of both the antigen and the antibody is the too! of choice to answer these questions. HEL was actually used as prototypic antigen because of the numerous natural variants available, and we have designed and expressed around .'iO mutants of HEL in the methylatropic yeast pichia pastoris. 'I hese mutants obtained bv replacing residues near or at the interface of HyHEL 1ÜTIEI. complex with Ala, will be used to study the interaction with HyHKL 10 and also with two related antibodies of close specificity. HyHKL H and HyHEL 2fi. The sensitivity <>f these related antibodies to the mutants will also shed light on the contribution of antibody residues in the interaction, by correlating the sensitivities to the primary sequence of these diidtibodies.

UR - http://www.scopus.com/inward/record.url?scp=33750197150&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33750197150&partnerID=8YFLogxK

M3 - Article

VL - 11

JO - FASEB Journal

JF - FASEB Journal

SN - 0892-6638

IS - 9

ER -