2018
DOI: 10.1080/09540105.2018.1440535
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Generation of panels of anti-idiotypic single-chain variable fragments mimicking Cry2Aa toxin using the chain shuffling technique

Abstract: Here, we firstly reported to generate panels of anti-idiotypic singlechain variable fragments (scFvs) of Bacillus thuringiensis Cry2Aa toxin by chain shuffling method. Light-and heavy-chain-shuffled libraries based on two parental clones were constructed. The signal of the light-chain-shuffled library was 4.2-fold higher than the heavy one. Solely after one round panning, eight mutants were rapidly isolated from the chain-shuffled libraries. The apparent affinity of the best clones was 2.65 × 10 6 M −1 , a 2.5… Show more

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Cited by 9 publications
(15 citation statements)
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“…The successful isolation of Ab2s from scFv libraries can be improved. For example, a study using an Ab1 for the protein toxin Cry2Aa rapidly isolated eight Ab2s from scFvs libraries that they had previously only isolated two clones from, after using a chain shuffling technique [37]. However, this is an extra stage and complicates the process, which can clearly be simplified by using naïve nanobody libraries.…”
Section: Alternative Libraries and Ab2smentioning
confidence: 99%
“…The successful isolation of Ab2s from scFv libraries can be improved. For example, a study using an Ab1 for the protein toxin Cry2Aa rapidly isolated eight Ab2s from scFvs libraries that they had previously only isolated two clones from, after using a chain shuffling technique [37]. However, this is an extra stage and complicates the process, which can clearly be simplified by using naïve nanobody libraries.…”
Section: Alternative Libraries and Ab2smentioning
confidence: 99%
“…Thus, in comparison with immunoassays based on traditional chemically synthesized antigens, novel developed detection methods employing biosynthetic antigens of VHHs display preferable cost advantages and product stability, so they are suitable for commercial production and widespread use. Accordingly, several anti-idiotypic VHHs (AI-VHHs), including those for mycotoxin zearalenone, fumonisin B 1 , aflatoxin B 1 , Ochratoxin A, deoxynivalenol (DON), and other toxins, have been developed as toxin surrogates in immunoassays (Hao et al, 2020;Liu et al, 2018;Qiu et al, 2016;Shu et al, 2019;Wang et al, 2013;Wang et al, 2016;Zhang et al, 2019). However, these determination methods for mycotoxin surveillance only avoid the application one of the chemically synthetized mycotoxin conjugates or mycotoxin detection standard.…”
Section: Introductionmentioning
confidence: 99%
“…Ab2 is the second antibody that recognizes different epitopes within the hypervariable region of the primary antibody (Jerne, 1974). Alphatype Ab2 (Ab2α) and betatype Ab2 (Ab2β) are the two main types of anti-idiotypic antibodie (Lan et al, 2017;Liu et al, 2018). Ab2α recognizes an epitope, which is not part of the binding site (paratope) of the primary antibody, involved in antigen recognition.…”
Section: Introductionmentioning
confidence: 99%