Strategy for affinity maturation of an antibody with high evolvability to (4-hydroxy-3-nitrophenyl) acetyl hapten

Furukawa, Koji; Shimizu, Takeyuki; Murakami, Akikazu; Kono, Ryo; Nakagawa, Masatoshi; Sagawa, T.; Yamato, Ichiro; Azuma, Takachika

doi:10.1016/j.molimm.2006.10.023

Cited by 19 publications

(7 citation statements)

References 42 publications

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“…Anti-NP mAbs were classified into four types according to the amino acid residues that are critical for affinity maturation of anti-NP IgG antibodies. Many antibodies established in this study belonged to the three types (Tyr95, Tyr95W33L and Gly95(H) types) that have already been described (20,21). However, some mAbs did not have tyrosine or glycine at position 95 of the V H gene and were sorted as the X95 type ( Fig.…”

Section: The Igm Mab Repertoire Was Different From the Igg Repertoirementioning

confidence: 79%

“…Because we did not analyze sequences from single cells, we could not gain sequence information on pairing light chains. Amino acid replacements in light chains contribute to affinity maturation (20), so that we could not speculate on the affinities of mutated V H genes from PCs. We chose V H genes without mutations, assuming that their partner light chains were in germline configuration.…”

Section: Unmutated and X95-type V H Genes Were Frequently Found In Sementioning

confidence: 99%

“…Tyr95-type antibodies can increase affinity quickly because a single amino acid substitution at position 33 (referred to as the W33L mutation) can raise affinity to NP ~10-fold (17)(18)(19), with added mutations causing a modest increase in affinity. Gly95(H)-type antibodies require multiple mutations in both heavy and light chains for an increase in affinity (20,21). Therefore, affinity maturation progresses slowly, but after accumulating multiple mutations, their affinity reaches ~1000-fold higher than those of germline antibodies.…”

Section: Introductionmentioning

confidence: 99%

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Low-affinity IgM antibodies lacking somatic hypermutations are produced in the secondary response of C57BL/6 mice to (4-hydroxy-3-nitrophenyl)acetyl hapten

Murakami

Moriyama

Osako-Kabasawa

et al. 2013

International Immunology

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Class-switched memory B cells, which are generated through the processes of somatic hypermutation (SHM) and affinity-based selection in germinal centers, contribute to the production of affinity-matured IgG antibodies in the secondary immune response. However, changes in the affinity of IgM antibodies during the immune response have not yet been studied, although IgM(+) memory B cells have been shown to be generated. In order to understand the relationship between IgM affinity and the recall immune response, we prepared hybridomas producing anti-(4-hydroxy-3-nitrophenyl)acetyl (NP) IgM antibodies from C57BL/6 mice and from activation-induced cytidine deaminase (AID)-deficient mice. Binding analysis by ELISA showed that mAbs obtained from the secondary immune response contained IgM mAbs with affinity lower than the affinity of mAbs obtained from the primary response. By analyzing sequences of the IgM genes of hybridomas and plasma cells, we found many unmutated VH genes. VH genes that had neither tyrosine nor glycine at position 95 were frequent. The repertoire change may correlate with the lower affinity of IgM antibodies in the secondary response. The sequence and affinity changes in IgM antibodies were shown to be independent of SHM by analyzing hybridomas from AID-deficient mice. A functional assay revealed a reciprocal relationship between affinity and complement-dependent hemolytic activity toward NP-conjugated sheep RBCs; IgM antibodies with lower affinities had higher hemolytic activity. These findings indicate that lower affinity IgM antibodies with enhanced complement activation function are produced in the secondary immune response.

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Section: The Igm Mab Repertoire Was Different From the Igg Repertoirementioning

confidence: 79%

Section: Unmutated and X95-type V H Genes Were Frequently Found In Sementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Low-affinity IgM antibodies lacking somatic hypermutations are produced in the secondary response of C57BL/6 mice to (4-hydroxy-3-nitrophenyl)acetyl hapten

Murakami

Moriyama

Osako-Kabasawa

et al. 2013

International Immunology

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“…To date, our knowledge of the underlying mechanisms in mammals has attained a considerable degree of sophistication, incorporating not only the temporal dynamics of somatic mutation [4,5], but also the relative interplay between kinetic vs. thermodynamic aspects of antigen binding [6,7], and the impact of cellular trafficking and differentiation [8,9].…”

Section: Introductionmentioning

confidence: 99%

The differential dynamics of antibody subpopulation expression during affinity maturation in a teleost

Kaattari

2011

Fish & Shellfish Immunology

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“…With the increasing number of antibody structures analysed, computer-aided design for antibody in vitro affinity maturation is becoming more reliable and convenient. This technology offers a significant advantage over other methods with greatly improved efficacy and success, because it can control the mutation sites within a certain range and target a few or even single amino acid sites 15 – 17 . Rodrigo Barderas et al .…”

Section: Introductionmentioning

confidence: 99%

A Highly Sensitive Detection System based on Proximity-dependent Hybridization with Computer-aided Affinity Maturation of a scFv Antibody

Wang

Liang

et al. 2018

Sci Rep

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The hepatitis B virus (HBV) infection is a critical health problem worldwide, and HBV preS1 is an important biomarker for monitoring HBV infection. Previously, we found that a murine monoclonal antibody, mAb-D8, targets the preS1 (aa91-107) fragment of HBV. To improve its performance, we prepared the single-chain variable region of mAb-D8 (scFvD8) and constructed the three-dimensional structure of the scFvD8-preS1 (aa91-107) complex by computer modelling. The affinity of scFvD8 was markedly increased by the introduction of mutations L96Tyr to Ser and H98Asp to Ser. Furthermore, a highly sensitive immunosensor was designed based on a proximity-dependent hybridization strategy in which the preS1 antigen competitively reacts with an antibody labelled with DNA, resulting in decreased proximity-dependent hybridization and increased electrochemical signal from the Fc fragment, which can be used for the quantisation of preS1. The results showed a wide detection range from 1 pM to 50 pM with a detection limit of 0.1 pM. The sensitivity and specificity of this immunosensor in clinical serum samples were 100% and 96%, respectively. This study provides a novel system based on proximity-dependent hybridization and the scFv antibody fragment for the rapid quantisation of antigens of interest with a high sensitivity.

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Strategy for affinity maturation of an antibody with high evolvability to (4-hydroxy-3-nitrophenyl) acetyl hapten

Cited by 19 publications

References 42 publications

Low-affinity IgM antibodies lacking somatic hypermutations are produced in the secondary response of C57BL/6 mice to (4-hydroxy-3-nitrophenyl)acetyl hapten

Low-affinity IgM antibodies lacking somatic hypermutations are produced in the secondary response of C57BL/6 mice to (4-hydroxy-3-nitrophenyl)acetyl hapten

The differential dynamics of antibody subpopulation expression during affinity maturation in a teleost

A Highly Sensitive Detection System based on Proximity-dependent Hybridization with Computer-aided Affinity Maturation of a scFv Antibody

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