Fahad Ahmed S, Gutiérrez-Gonzalez Matías F, Madan Bharat, DeKosky Brandon J
The Ragon Institute of Massachusetts General Hospital, Massachusetts Institute of Technology, and Harvard University, Cambridge, Massachusetts 02139, USA.
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Cold Spring Harb Protoc. 2025 Jan 2;2025(1):pdb.prot108626. doi: 10.1101/pdb.prot108626.
In vitro antibody evolution is a powerful technique for improving monoclonal antibodies. This can be achieved by generating artificial diversity on an antibody template, which can be done using different in vitro diversification techniques. The resulting libraries consist of single- or multimutant variants of a defined antibody template that are screened for improved function using antibody display. Here, we describe a bioinformatic protocol for tracking synthetic antibody variants using high-throughput sequencing across screening rounds, enabling efficient high-throughput interpretation of the function of individual mutations in sorted antibody display libraries. The protocol enables a user to achieve precision analysis and interpretation of clonal antibody variant data for discovery purposes, especially for high-throughput antibody engineering or optimization against target antigens.
体外抗体进化是一种用于改进单克隆抗体的强大技术。这可以通过在抗体模板上产生人工多样性来实现，这可以使用不同的体外多样化技术来完成。所得文库由定义的抗体模板的单突变或多突变变体组成，这些变体通过抗体展示筛选以获得改进的功能。在这里，我们描述了一种生物信息学方案，用于在筛选轮次中使用高通量测序跟踪合成抗体变体，从而能够对分选的抗体展示文库中单个突变的功能进行高效的高通量解读。该方案使用户能够为发现目的实现对克隆抗体变体数据的精确分析和解读，特别是对于针对靶抗原的高通量抗体工程或优化。
