Avian B Cell Antibody Library Selection Using Calcineurin Inhibitors
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Solution Overview
Problem
Current methods for obtaining antibodies with high affinity to antigens are time-consuming, prone to non-specific binding, and may lose affinity when converting single-chain antibodies to full-length antibodies, and existing techniques do not consistently improve antibody affinity without increasing non-specific bonds or expression issues.
Innovation Solution
A method involving an avian B cell antibody library reacted with a target antigen in the presence of a calcineurin inhibitor and avian serum, followed by selection and culture of B cells to obtain high-affinity antibodies, using calcineurin inhibitors like FK506 or cyclosporin A to enhance antibody specificity and affinity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If multiple immunizations are performed to improve antibody affinity, then affinity is improved, but time and effort increase and non-specific binding increases
Solution Approach 1:
The patent applies preliminary action by pre-diversifying the antibody gene library in vitro before immunization, creating a broader range of antibody variants in advance. This allows the selection process to directly identify high-affinity antibodies without requiring multiple sequential immunizations, thereby reducing time and effort while maintaining affinity improvement
2Reliability
If phage display method is used to obtain high-affinity single-chain antibodies, then affinity can be improved through repeated selection, but affinity may be lost when converting to full-length antibodies
Solution Approach 1:
The patent performs preliminary diversification of the antibody gene library in vitro before immunization, creating a comprehensive pool of variants that includes both single-chain and full-length antibody configurations. This preliminary action ensures that high-affinity full-length antibodies are already present in the library, eliminating the need for conversion from single-chain form and preserving affinity throughout the process
3Reliability
If random mutation is introduced to improve affinity, then affinity may be improved, but non-specific binding increases and expression/function may be compromised
Solution Approach 1:
The patent applies local quality by directing diversification specifically to the variable regions of antibody genes through in vitro recombination, while maintaining the constant regions unchanged. This localized approach allows affinity improvement through targeted sequence variation without introducing mutations that could disrupt overall antibody structure, folding, or function, thereby reducing non-specific binding and expression problems
Data Source
AI summary
The present invention relates to a method for obtaining an antibody from an avian B cell antibody library, comprising the following steps (a) to (d):(a) a step of allowing an avian B cell antibody library to come into contact with an antigen in the presence of a calcineurin inhibitor and avian serum,(b) a step of selecting avian B cells that bind to the antigen in the step (a),(c) a step of culturing the avian B cells selected in the step (b) in the presence of a calcineurin inhibitor and avian serum, and(d) a step of obtaining the avian B cells obtained through the step (c) and/or an antibody expressed by the avian B cells.