What is a phage or bacteriophage?
Phages are viruses that infect bacterial cells. There are several types of phages that have been used as vehicles for phage display including Ff filamentous phage, Lambda and T7. In phage display, the bacteriophages are used to construct different phage libraries as peptide libraries or antibody library
History of phage display
Phage display technology was first introduced in 1985 by George Smith. After that many advantages have been introduced including the way to construct a phage library like antibody library to select phage display antibodies.
Advantages of phage display
Phage display is a system for large scale study and selection of proteins, peptide and antibodies based on their binding affinity and specificity. One advantage of phage display is the enormous diversity of variant proteins that can be represented in a phage library. Phage display provides a means of rapidly screening large numbers of proteins against potential binding partners.
humanized monoclonal

Humanized monoclonal antibody

Humanized monoclonal antibody overview

 

“How can be produced a humanized monoclonal antibody ?”  by humanization of antibodies, the antibodies from non-human species whose amino acid sequences are changed to improve their similarity to antibody variants produced naturally in humans. Before rodent antibodies can be used in clinical application is essential to humanize them to decrease their immunogenicity and increase their activity in the human immune system.

 

Humanized monoclonal antibody methodology

Due the advances in antibody engineering and molecular biology, it is possible to use several different methods to obtain a humanized monoclonal antibody, including CDR grafting and screening of a human phage display antibody library.

 

Humanized monoclonal antibody production by CDR grafting technique

Production of humanized monoclonal antibody  by the CDR grafting technique consists in transferring the antigen-binding loops of the non-human antibody to an artificially chosen human framework.

When it is applying the CDR grafting technique to obtain a humanized monoclonal antibody, it should be taken in consideration several factors, for example the native loop conformations should be preserved as long it is possible to conserve the antibody binding affinity and specificity, the immunogenic sequences should be removed, and the antibody effector functions should be conserved.

 

Humanized monoclonal antibody production by selecting antibodies from a human antibody library

Human antibody libraries can be made isolating from the VH and VL repertoire from b-cells from healthly human donors. The VH and VL are put together as Fab or scFV to be expressed on the surface of phages in fussion with the pIII protein. The antibody repertoire is enough high (size of 10e9 -10e10) to isolate antibodies against almost every antigen. The screening procedure consists in different steps of binding, whasing, and elutions named panning or biopanning. Phage carrying antibodies againts the target protein are isolated and the Fab and scFv fragment are expressed in E.coli or mammalian expresssion systems.

 

Humanized monoclonal antibody production services

Vrisko Limited offers production of humanized monoclonal antibody production services  including CDR grafting and screening of a human phage display antibody library. Send us an inquiry for more information of our humanized monoclonal antibody production services. Online inquiry

 

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