Anti-Trop-2 Antibody Screening for Native Binding and Internalization
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing anti-Trop-2 antibodies exhibit insufficient affinity for the native conformation of Trop-2 extracellular domain in vivo, leading to poor tumor targeting activity and cytotoxin delivery capacity, which hinders their effectiveness in clinical trials.
Innovation Solution
A method involving hybridoma screening and humanization technologies to develop anti-Trop-2 monoclonal antibodies with high affinity for the native conformation of human Trop-2 extracellular domain, using recombinant Trop-2 protein as an immunogen, followed by ELISA screening and rescreening with Trop-2 positive cells on the cell membrane surface to ensure specificity and binding ability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If high affinity antibodies against Trop-2 are screened in vitro using recombinant proteins, then antibody affinity to recombinant Trop-2 is improved, but tumor targeting activity and cytotoxin delivering capacity in vivo deteriorate
Solution Approach 1:
The patent uses recombinant Trop-2 protein as a copy of the native Trop-2 extracellular domain for immunization and screening. This copy preserves the conformational epitopes of the native protein, enabling the generation of antibodies that recognize both the recombinant and native forms with high affinity, thereby resolving the discrepancy between in vitro affinity and in vivo targeting activity
Solution Approach 2:
The patent changes the screening parameter from recognizing denatured or linear epitopes to recognizing conformational epitopes on the surface of recombinant Trop-2 protein. This parameter change ensures that antibodies selected in vitro maintain their binding capability to the native Trop-2 extracellular domain in vivo, improving both affinity and targeting activity
2Ease of operation
If conventional antibody screening methods are used, then screening process is simplified, but antibody internalization ability deteriorates
Solution Approach 1:
The patent performs preliminary selection of hybridoma clones that bind to conformational epitopes on recombinant Trop-2 protein before final screening. This preliminary action ensures that only antibodies with the correct epitope recognition are advanced to internalization assays, efficiently enriching for candidates with both binding affinity and internalization capability
Solution Approach 2:
The patent uses recombinant Trop-2 protein as an intermediary between the hybridoma screening and the assessment of internalization ability. This intermediary serves as a bridge that connects antibody binding to the native conformation with functional internalization, enabling the selection of antibodies that possess both properties without requiring complex direct assays
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 developed antibodies demonstrate high specificity and internalization ability, making them suitable for ADC drugs and improving preclinical performance closer to in vivo results.
Implementation Method 1
The developed antibodies demonstrate high specificity and internalization ability, making them suitable for ADC drugs
Implementation Method 2
The developed antibodies demonstrate high specificity and internalization ability
Data Source
AI summary
The present invention provides an antibody binding a human tumor-associated calcium signal sensor 2 (Trop-2) protein or fragments thereof, and use of the antibody or fragments thereof in preventing or treating diseases. The antibody or fragments thereof of the present invention can effectively bind to the human Trop-2 protein, and have internalization activity, and the internalization activity is enhanced after ADC drug labeling, and the in vivo efficacy and safety of a mouse model are not lower than those of a control antibody.