Recombinant Antibodies and Non-Antibody Scaffolds for Immunoassays

Ban, Bhupal; Blake, Diane A.

doi:10.5772/35983

Cited by 2 publications

(2 citation statements)

References 88 publications

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“…Currently, monoclonal antibodies (mAbs) are key biologics for several applications such as basic science research, therapeutics protein, and clinical diagnostics assays such as immunoassays, immunohistochemistry, flow cytometry, and immuno-PET scanning. In addition, antibodies are used in proteome research for the identification of biomarkers [1]. Advancements in the area of immunoinformatic and recombinant antibodies have changed the applications of domain antibodies such as the single chain fragment variable (scFv), the antigen binding fragment (Fab), and single domain of variable heavy homodimers (VHH) are very popular reagents for basic scientific and clinical applications [2].…”

Section: Introductionmentioning

confidence: 99%

Optimization of Methods for the Production and Refolding of Biologically Active Disulfide Bond-Rich Antibody Fragments in Microbial Hosts

2020

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Antibodies have been used for basic research, clinical diagnostics, and therapeutic applications. Escherichia coli is one of the organisms of choice for the production of recombinant antibodies. Variable antibody genes have canonical and non-canonical disulfide bonds that are formed by the oxidation of a pair of cysteines. However, the high-level expression of an antibody is an inherent problem to the process of disulfide bond formation, ultimately leading to mispairing of cysteines which can cause misfolding and aggregation as inclusion bodies (IBs). This study demonstrated that fragment antibodies are either secreted to the periplasm as soluble proteins or expressed in the cytoplasm as insoluble inclusion bodies when expressed using engineered bacterial host strains with optimal culture conditions. It was observed that moderate-solubilization and an in vitro matrix that associated refolding strategies with redox pairing more correctly folded, structured, and yielded functionally active antibody fragments than the one achieved by a direct dilution method in the absence of a redox pair. However, natural antibodies have canonical and non-canonical disulfide bonds that need a more elaborate refolding process in the presence of optimal concentrations of chaotropic denaturants and redox agents to obtain correctly folded disulfide bonds and high yield antibodies that retain biological activity.

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Section: Introductionmentioning

confidence: 99%

Optimization of Methods for the Production and Refolding of Biologically Active Disulfide Bond-Rich Antibody Fragments in Microbial Hosts

2020

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“…Одним з перспективних напрямків у галузі розвитку сенсорних пристроїв, які об'єднують специфічність дії антитіл і ферментів, є пошук альтернативи природним антитілам. Це пов'язано з тим, що антитілам притаманні такі недоліки, як [3]:…”

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Synthetic Anti-Melamine Receptor Based on Molecularly Imprinted Polymers for Pseudo-Elisa

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2015

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Показана можливість застосування синтетичних рецепторів (отриманих на базі технології твердофазного молекулярного імпринтингу) як дешевої альтернативи ан-титілам в ході проведення твердофазного імуноферментного визначення меламіну. Попе-редні квантовохімічні розрахунки дали можливість обґрунтувати вибір найкращого кан-дидата-мономера (серед міні-бібліотеки з 13 мономерів) під час створення за технологією молекулярного імпринтингу «штучного рецептора» на меламін, а саме, акриламідо-2-метил-1-пропансульфонової кислоти. Межа визначення меламіну конкурентним методом становила 0.0058 нM (5.8 пМ), лінійний діапазон концентрацій: 0.01-1 нM. Чутливість визначення меламіну даним методом вища (втричі), ніж ELISA-аналіз, що базується на антитілах. Ключові слова: молекулярно імпринтований полімер, наночастинки, твердофазний імуноферментний аналіз, штучний рецептор, меламін, конкурентний аналіз

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Recombinant Antibodies and Non-Antibody Scaffolds for Immunoassays

Cited by 2 publications

References 88 publications

Optimization of Methods for the Production and Refolding of Biologically Active Disulfide Bond-Rich Antibody Fragments in Microbial Hosts

Optimization of Methods for the Production and Refolding of Biologically Active Disulfide Bond-Rich Antibody Fragments in Microbial Hosts

Synthetic Anti-Melamine Receptor Based on Molecularly Imprinted Polymers for Pseudo-Elisa

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