Developing Recombinant Antibodies by Phage Display Against Infectious Diseases and Toxins for Diagnostics and Therapy

Abstract

Antibodies are essential molecules for diagnosis and treatment of diseases caused by pathogens and their toxins. Antibodies were integrated in our medical repertoire against infectious diseases more than hundred years ago by using animal sera to treat tetanus and diphtheria. In these days, most developed therapeutic antibodies target cancer or autoimmune diseases. The COVID-19 pandemic was a reminder about the importance of antibodies for therapy against infectious diseases. While monoclonal antibodies could be generated by hybridoma technology since the 70ies of the former century, nowadays antibody phage display, among other display technologies, is robustly established to discover new human monoclonal antibodies. Phage display is an in vitro technology which confers the potential for generating antibodies from universal libraries against any conceivable molecule of sufficient size and omits the limitations of the immune systems. If convalescent patients or immunized/infected animals are available, it is possible to construct immune phage display libraries to select in vivo affinity-matured antibodies. A further advantage is the availability of the DNA sequence encoding the phage displayed antibody fragment, which is packaged in the phage particles. Therefore, the selected antibody fragments can be rapidly further engineered in any needed antibody format according to the requirements of the final application. In this review, we present an overview of phage display derived recombinant antibodies against bacterial, viral and eukaryotic pathogens, as well as microbial toxins, intended for diagnostic and therapeutic applications.

Phage Display-Derived Monoclonal Antibodies Against Internalins A and B Allow Specific Detection of Listeria monocytogenes

“…Another possibility may be due to a low number of human antibody germline sequences against this antigen. Even though it is unlikely that phage display does not provide antibodies against bacterial antigens ( 33 ), previous works described regions of bacterial antigens, including Listeria spp., that provide few antibodies ( 23 ). This indicates that InlB may be substantially different from InlA (identity = 28%), which leads to low number of hits when performing panning.…”

Improving antibody affinity through <i>in vitro</i> mutagenesis in complementarity determining regions

“…Phage display is a technology established for generating human monoclonal antibodies. This approach is an in vitro technology that confers the potential for generating antibodies from phage libraries against any conceivable molecule of sufficient size while overcoming limitations of in vivo immunization [ 5 , 7 ] . With full-length purified GPC3 and the RBD domain of SARS-CoV-2 as antigens, we screened the Tomlinson I and J libraries and obtained specific antibody fragments 42A1 and I4A3, from each respectively.…”

“…Phage display is a widely used and is a powerful technology that allows the display of antibody fragments on the surface of filamentous bacteriophages infecting E. coli [ 4 ] . This approach uses an in vitro selection process that does not have to rely on immunization, and can make use of entirely human gene repertoires [ 5 – 7 ] .…”

Can the SARS-CoV-2 Omicron Variant Confer Natural Immunity against COVID-19?

“…It occurs when a disease-causing organism stimulates the immune system to produce antibodies against that disease [ 29 ]. Antibodies are proteins that the body produces specifically to neutralize or destroy pathogenic organisms, their toxins, and fragments thereof [ 37 ]. Vaccine-induced immunity is acquired by vaccination with a killed or weakened form of the disease-causing organism [ 38 , 39 ].…”