High-throughput generation and engineering of recombinant human antibodies

Area of Science:

• Biotechnology
• Immunology
• Molecular Biology

Background:

• The Human Combinatorial Antibody Library (HuCAL) is a single-chain fragment variable (scFv)-based phage display library.
• It comprises 2x10^9 members optimized for high-throughput generation and engineering of human antibodies.

Purpose of the Study:

• To demonstrate the feasibility of automated, miniaturized screening of the HuCAL-scFv library.
• To highlight the library's modular design for converting scFv into various antibody formats and fusions.
• To showcase HuCAL as a convenient source for diverse human antibody applications.

Main Methods:

• Utilizing a single-chain Fv-based phage display library (HuCAL-scFv) with 2x10^9 members.
• Expressing antibodies in high levels in Escherichia coli for automated panning and screening.
• Employing a modular design for converting scFv into Fab, miniantibody, and immunoglobulin formats.

Main Results:

• 61% of the library genes code for functional scFv antibodies.
• High-level expression in E. coli enables automated, miniaturized screening (96- and 384-well formats).
• Facilitated conversion into multiple formats and fusion with effector functions/tags.

Conclusions:

• The HuCAL principle enables rapid, high-throughput development of human antibodies.
• HuCAL-scFv is suitable for automated screening and engineering.
• The library's modularity supports diverse therapeutic and research applications.