Anti-HER2 Antibody CDR Optimization for High-Sensitivity Detection
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Solution Overview
Problem
Current methods for detecting HER2 in biological samples are not sufficiently effective, and existing anti-HER2 antibodies lack specificity and sensitivity, particularly in diagnosing HER-positive tumors.
Innovation Solution
Development of a novel anti-HER2 antibody with specific heavy and light chain variable regions and a nucleic acid molecule encoding these regions, used in an immunoenzymatic ELISA assay for sensitive and specific detection of HER2 in biological samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If existing anti-HER2 antibodies are used for detection, then the detection method can be implemented, but the sensitivity and specificity are insufficient
Solution Approach 1:
The patent develops novel anti-HER2 antibodies with optimized amino acid sequences in the variable regions, specifically modifying the complementarity determining regions (CDRs) to enhance binding affinity and specificity. This parameter change at the molecular level directly improves detection sensitivity to 0.5 ng/ml threshold and eliminates cross-reactivity with HER2-negative samples, thereby resolving the contradiction between measurement precision and reliability
Solution Approach 2:
The patent creates recombinant antibodies that replicate and optimize the binding properties of natural antibodies. By copying and refining the variable region sequences through recombinant DNA technology, the invention produces antibodies with enhanced specificity and sensitivity that overcome the limitations of existing detection methods while maintaining the fundamental antibody-antigen recognition mechanism
2Measurement precision
If conventional detection methods are used, then the procedure can be completed, but the sensitivity threshold is too high (cannot detect low concentrations)
Solution Approach 1:
The patent achieves a detection sensitivity threshold of 0.5 ng/ml by optimizing antibody concentration, incubation time, and washing conditions in the ELISA protocol. The standardized assay development process, despite initial complexity, establishes a robust framework that can be replicated across laboratories, ultimately simplifying implementation while achieving superior sensitivity compared to conventional methods
3Reliability
If existing antibodies are used, then the current diagnostic workflow can be maintained, but cross-reactivity with HER2-negative samples occurs
Solution Approach 1:
The patent introduces strict specificity criteria for antibody selection, focusing on localized modifications to the variable region sequences, particularly the CDRs. This localized optimization ensures high specificity for HER2-positive samples while eliminating cross-reactivity with HER2-negative samples, thereby improving reliability without requiring complex multi-antibody panels or additional diagnostic steps
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The novel anti-HER2 antibody provides high antigen binding strength and specificity, enabling detection of HER2 with a low sensitivity threshold of 0.5 ng/ml, suitable for quantifying HER2 within the standard curve range of 15.6-100 ng/ml, and allows for the determination of higher concentrations through dilutions, facilitating accurate diagnosis and therapeutic applications.
Implementation Method 1
The novel anti-HER2 antibody provides high antigen binding strength and specificity, enabling detection of HER2
Implementation Method 2
immuno-enzymatic ELISA assay for sensitive and specific detection of HER2
Data Source
Figure 1~2B
Figure 3A~3C
Figure 4A~4C
AI summary
Invention relates to an anti-HER2 antibody, characterised in that it comprises: a) a heavy chain variable region comprising CDR1, CDR2 and CDR3, in which CDR1 is an amino acid sequence of GYSFTGYN (SEQ ID No. 1) or GYSFTDYN (SEQ ID No. 2), CDR2 is an amino acid sequence of IDPNYGT (SEQ ID No. 3) or IDHYNGNT (SEQ ID No. 4), CDR3 is an amino acid sequence of ARRKVNYEAWFAF (SEQ ID No. 5) or ARHYDYGAMDY (SEQ ID No. 6); b) a light chain variable region comprising CDR1, CDR2 and CDR3 in which CDR1 is an amino acid sequence of ESVDSYGNSF (SEQ ID No. 7) or QNVGTN (SEQ ID No. 8), CDR2 is an amino acid sequence of RAS (SEQ ID No. 9) or SAS (SEQ ID No. 10), CDR3 constitutes an amino acid sequence of QQSNEDPFT (SEQ ID No. 11) or QHYYIYPLT (SEQ ID No. 12). Invention further relates to a nucleic acid molecule encoding the variable region of an antibody according to invention or a fragment thereof. Invention further relates to a method of detecting HER2 in a biological sample, comprising contacting the sample with an HER2-binding protein and detecting a complex, wherein the detection of said complex indicates HER2 expression in the sample, characterised in that HER2-binding protein is the antibody according to invention. Invention further relates to an immuno-enzymatic ELISA assay for the detection of HER2 antigen, characterised in that at least one antibody according to invention is used for the detection of antigen. Invention further relates to the use of antibody according to invention for producing antibody-drug conjugates and to the use of SEQ ID No. 17-28 nucleotide sequences encoding variable regions of the antibodies according to invention for the CAR-T production.