Thermodynamic consequences of grafting enhanced affinity toward the mutated antigen onto an antibody. The case of anti-lysozyme antibody HyHEL-10

Yoshiyuki Nishimiya, Kouhei Tsumoto, Mitsunori Shiroishi, Katsuhide Yutani, Izumi Kumagai

研究成果: ジャーナルへの寄稿記事

21 引用 (Scopus)

抄録

In order to address the mechanism of enhancement of the affinity of an antibody toward an antigen from a thermodynamic viewpoint, anti-hen lysozyme (HEL) antibody HyHEL-10, which also recognize the mutated antigen turkey lysozyme (TEL) with reduced affinity, was examined. Grafting high affinity toward TEL onto HyHEL-10 was performed by saturation mutagenesis into four residues (Tyr53, Ser54, Ser56, and Tyr58) in complementarity- determining region 2 of the heavy chain (CDR-H2) followed by selection with affinity for TEL. Several clones enriched have a Phe residue at site 58. Thermodynamic analyses showed that the clones selected had experienced a greater than 3-fold affinity increase toward TEL in comparison with wild-type Fv, originating from an increase in negative enthalpy change. Substitution of HyHEL-10 HTyr58 with Phe led to the increase in negative enthalpy change and to almost identical affinity for TEL in comparison with mutants selected, indicating that mutations at other sites decrease the entropy loss despite little contribution to the affinity for TEL. These results suggest that the affinity of an antibody toward the antigen is enhanced by the increase in enthalpy change by some limited mutation, and excess entropy loss due to the mutation is decreased by other energetically neutral mutations.

元の言語英語
ページ(範囲)12813-12820
ページ数8
ジャーナルJournal of Biological Chemistry
275
発行部数17
DOI
出版物ステータス出版済み - 4 28 2000

Fingerprint

Muramidase
Thermodynamics
Anti-Idiotypic Antibodies
Antigens
Antibodies
Enthalpy
Mutation
Antibody Affinity
Entropy
Clone Cells
Complementarity Determining Regions
Mutagenesis
Substitution reactions

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

これを引用

Thermodynamic consequences of grafting enhanced affinity toward the mutated antigen onto an antibody. The case of anti-lysozyme antibody HyHEL-10. / Nishimiya, Yoshiyuki; Tsumoto, Kouhei; Shiroishi, Mitsunori; Yutani, Katsuhide; Kumagai, Izumi.

:: Journal of Biological Chemistry, 巻 275, 番号 17, 28.04.2000, p. 12813-12820.

研究成果: ジャーナルへの寄稿記事

Nishimiya, Yoshiyuki ; Tsumoto, Kouhei ; Shiroishi, Mitsunori ; Yutani, Katsuhide ; Kumagai, Izumi. / Thermodynamic consequences of grafting enhanced affinity toward the mutated antigen onto an antibody. The case of anti-lysozyme antibody HyHEL-10. :: Journal of Biological Chemistry. 2000 ; 巻 275, 番号 17. pp. 12813-12820.
@article{e366c0929cb24b8093432d196086f560,
title = "Thermodynamic consequences of grafting enhanced affinity toward the mutated antigen onto an antibody. The case of anti-lysozyme antibody HyHEL-10",
abstract = "In order to address the mechanism of enhancement of the affinity of an antibody toward an antigen from a thermodynamic viewpoint, anti-hen lysozyme (HEL) antibody HyHEL-10, which also recognize the mutated antigen turkey lysozyme (TEL) with reduced affinity, was examined. Grafting high affinity toward TEL onto HyHEL-10 was performed by saturation mutagenesis into four residues (Tyr53, Ser54, Ser56, and Tyr58) in complementarity- determining region 2 of the heavy chain (CDR-H2) followed by selection with affinity for TEL. Several clones enriched have a Phe residue at site 58. Thermodynamic analyses showed that the clones selected had experienced a greater than 3-fold affinity increase toward TEL in comparison with wild-type Fv, originating from an increase in negative enthalpy change. Substitution of HyHEL-10 HTyr58 with Phe led to the increase in negative enthalpy change and to almost identical affinity for TEL in comparison with mutants selected, indicating that mutations at other sites decrease the entropy loss despite little contribution to the affinity for TEL. These results suggest that the affinity of an antibody toward the antigen is enhanced by the increase in enthalpy change by some limited mutation, and excess entropy loss due to the mutation is decreased by other energetically neutral mutations.",
author = "Yoshiyuki Nishimiya and Kouhei Tsumoto and Mitsunori Shiroishi and Katsuhide Yutani and Izumi Kumagai",
year = "2000",
month = "4",
day = "28",
doi = "10.1074/jbc.275.17.12813",
language = "English",
volume = "275",
pages = "12813--12820",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "17",

}

TY - JOUR

T1 - Thermodynamic consequences of grafting enhanced affinity toward the mutated antigen onto an antibody. The case of anti-lysozyme antibody HyHEL-10

AU - Nishimiya, Yoshiyuki

AU - Tsumoto, Kouhei

AU - Shiroishi, Mitsunori

AU - Yutani, Katsuhide

AU - Kumagai, Izumi

PY - 2000/4/28

Y1 - 2000/4/28

N2 - In order to address the mechanism of enhancement of the affinity of an antibody toward an antigen from a thermodynamic viewpoint, anti-hen lysozyme (HEL) antibody HyHEL-10, which also recognize the mutated antigen turkey lysozyme (TEL) with reduced affinity, was examined. Grafting high affinity toward TEL onto HyHEL-10 was performed by saturation mutagenesis into four residues (Tyr53, Ser54, Ser56, and Tyr58) in complementarity- determining region 2 of the heavy chain (CDR-H2) followed by selection with affinity for TEL. Several clones enriched have a Phe residue at site 58. Thermodynamic analyses showed that the clones selected had experienced a greater than 3-fold affinity increase toward TEL in comparison with wild-type Fv, originating from an increase in negative enthalpy change. Substitution of HyHEL-10 HTyr58 with Phe led to the increase in negative enthalpy change and to almost identical affinity for TEL in comparison with mutants selected, indicating that mutations at other sites decrease the entropy loss despite little contribution to the affinity for TEL. These results suggest that the affinity of an antibody toward the antigen is enhanced by the increase in enthalpy change by some limited mutation, and excess entropy loss due to the mutation is decreased by other energetically neutral mutations.

AB - In order to address the mechanism of enhancement of the affinity of an antibody toward an antigen from a thermodynamic viewpoint, anti-hen lysozyme (HEL) antibody HyHEL-10, which also recognize the mutated antigen turkey lysozyme (TEL) with reduced affinity, was examined. Grafting high affinity toward TEL onto HyHEL-10 was performed by saturation mutagenesis into four residues (Tyr53, Ser54, Ser56, and Tyr58) in complementarity- determining region 2 of the heavy chain (CDR-H2) followed by selection with affinity for TEL. Several clones enriched have a Phe residue at site 58. Thermodynamic analyses showed that the clones selected had experienced a greater than 3-fold affinity increase toward TEL in comparison with wild-type Fv, originating from an increase in negative enthalpy change. Substitution of HyHEL-10 HTyr58 with Phe led to the increase in negative enthalpy change and to almost identical affinity for TEL in comparison with mutants selected, indicating that mutations at other sites decrease the entropy loss despite little contribution to the affinity for TEL. These results suggest that the affinity of an antibody toward the antigen is enhanced by the increase in enthalpy change by some limited mutation, and excess entropy loss due to the mutation is decreased by other energetically neutral mutations.

UR - http://www.scopus.com/inward/record.url?scp=0034724710&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034724710&partnerID=8YFLogxK

U2 - 10.1074/jbc.275.17.12813

DO - 10.1074/jbc.275.17.12813

M3 - Article

C2 - 10777579

AN - SCOPUS:0034724710

VL - 275

SP - 12813

EP - 12820

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 17

ER -