Optimization of stereospecific targeting technique for selective production of monoclonal antibodies against native ephrin type-A receptor 2

Yamasaki, Yasuhiro; Miyamae, Chiho; Isozaki, Yushi; Ichikawa, Keisuke; Kaneko, Yoshiki; Oda, Yasuyuki; Murayama, Takashi; Sakurai, Takashi; Tamai, Katsuyuki; Tsumoto, K.; Tomita, Masahiro

doi:10.1016/j.jim.2020.112813

Cited by 3 publications

(10 citation statements)

References 20 publications

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“…( 3 ) developed a stereospecific targeting (SST) method where myeloma cells selectively fuse with B cells that produce antigen-specific antibodies prior to the cell fusion. A recent study reported an optimized version of this method, which has improved the efficiency of obtaining hybridomas that produce the target antibody ( 11 ).…”

Section: Discussionmentioning

confidence: 99%

“…A method of screening clones that produce antigen-specific antibodies after cell fusion is an FCM-based methodology that enables the selection of a single cell from a bulk mixture of fused hybridoma cells. The method does not require special reagents, is easy to set up, and can be applied in many different types of laboratories ( 8 , 11 , 12 , 13 , 14 ). By advancing both methods (SST and MIHS), it will be possible to obtain a variety of monoclonal antibodies suitable for specific applications.…”

Section: Discussionmentioning

confidence: 99%

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Rapid and reliable hybridoma screening method that is suitable for production of functional structure-recognizing monoclonal antibody

Sakaguchi

Nakajima

Sawano

et al. 2021

Journal of Bioscience and Bioengineering

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Monoclonal antibodies are extremely valuable functional biomaterials that are widely used not only in life science research but also in antibody drugs and test drugs. There is also a strong need to develop high-quality neutralizing antibodies as soon as possible in order to stop the rapid spread of new infectious diseases such as the SARS-CoV-2 virus. This study has developed a membrane-type immunoglobulin-directed hybridoma screening (MIHS) method for obtaining high-quality monoclonal antibodies with high efficiency and high speed. In addition to these advantages, this paper demonstrates that the MIHS method can selectively obtain monoclonal antibodies that specifically recognize the functional structure of proteins. The MIHS method is a useful technology that greatly contributes to the research community because it can be easily introduced in any laboratory that uses a flow cytometer.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Rapid and reliable hybridoma screening method that is suitable for production of functional structure-recognizing monoclonal antibody

Sakaguchi

Nakajima

Sawano

et al. 2021

Journal of Bioscience and Bioengineering

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“…The idea is to promote a further stage in the B cell maturation. Indeed, an increase in serum antibody titers, when compared to the results of gene immunization only, could be observed ( Table 1 ) ( Yamasaki et al., 2020 ). The second step involves the preselection of conformational epitope-recognizing B cells.…”

Section: Hybridoma Technologymentioning

confidence: 96%

“…The screening of hybridomas secreting the desired mAbs, the fourth step, makes use of the native antigen targets expressed on a cell surface. The clone selection may include an additional step to discard the undesirable clones by using recombinant protein, which may contain partially denatured structures ( Yamasaki et al., 2020 ). The SST method provides more than 50% positivity for B lymphocyte-myeloma cell fusion, and more than 24% of the generated clones were found to secrete the desired mAbs ( Yamasaki et al., 2020 ).…”

Section: Hybridoma Technologymentioning

confidence: 99%

Hybridoma technology: is it still useful?

Moraes

Hamaguchi

Braggion

et al. 2021

Current Research in Immunology

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“…We previously reported a novel technology termed optimized stereospecific targeting (SST) [8][9][10], which consists of three important steps: (1) DNA immunization;…”

Section: Introductionmentioning

confidence: 99%

Class-Switching of B Lymphocytes by DNA and Cell Immunization for Stereospecific Monoclonal Antibodies against Native GPCR

Isozaki

Tsumoto

Tomita

2021

Immuno

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To develop efficient applications of monoclonal antibodies for therapeutic purposes, stereospecific recognition of the target antigens is needed. DNA immunization is one of the best methods for sensitizing B lymphocytes that can produce conformation-specific antibodies. Here we verified the class-switching of monoclonal antibodies by DNA immunization followed by cell immunization for the generation of stereospecific monoclonal antibodies against native G protein-coupled receptor (GPCR) using the optimized stereospecific targeting (SST) technique. This technology facilitates the efficient selection of sensitized B lymphocytes through specific interaction with the intact antigen via B-cell receptors (BCRs). We demonstrate that multiple DNA immunizations followed by a single cell immunization in combination with a longer sensitization period (three to four months) are an appropriate sensitizing strategy for the generation of IgG-type stereospecific monoclonal antibodies by class-switching, and the characteristics of antibody production could be transferred to hybridoma cells provided by the optimized SST technique.

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Optimization of stereospecific targeting technique for selective production of monoclonal antibodies against native ephrin type-A receptor 2

Cited by 3 publications

References 20 publications

Rapid and reliable hybridoma screening method that is suitable for production of functional structure-recognizing monoclonal antibody

Rapid and reliable hybridoma screening method that is suitable for production of functional structure-recognizing monoclonal antibody

Hybridoma technology: is it still useful?

Class-Switching of B Lymphocytes by DNA and Cell Immunization for Stereospecific Monoclonal Antibodies against Native GPCR

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