Protocol for isolating antigen-specific monoclonal antibodies from immunized mice utilizing the Beacon platform.

Area of Science:

• Immunology
• Molecular Biology
• Biotechnology

Background:

• Isolating antigen-specific plasma B cells has historically been difficult.
• The Beacon platform offers a novel solution using light-sorting technology.
• High-throughput screening of single cells for specific antigen recognition is now feasible.

Purpose of the Study:

• To present a detailed protocol for isolating antigen-specific plasma B cells.
• To demonstrate the application of the Beacon platform for B cell receptor (BCR) sequencing.
• To outline the subsequent cloning and expression of antibodies derived from these B cells.

Main Methods:

• Utilized the Beacon platform for high-throughput, single-cell sorting based on antigen specificity.
• Employed sequencing techniques to determine the B cell receptors (BCRs) from isolated plasma B cells.
• Performed molecular cloning and expression of antibodies encoded by the identified BCRs.

Main Results:

• Successfully isolated antigen-specific plasma B cells from immunized mouse models.
• Obtained sequences for B cell receptors (BCRs) from the sorted single cells.
• Expressed functional antibodies derived from the isolated and sequenced B cells.

Conclusions:

• The Beacon platform provides an effective method for isolating antigen-specific plasma B cells.
• This protocol facilitates the discovery and development of novel antibodies.
• The methodology is adaptable for use with human samples, broadening its potential applications.