Fab Antibody Humanization Service

Introduction of Fab Antibody Humanization ServiceFab

Human mouse chimeric mab with tumor nucleus (chTNT) is a novel antibody against tumor necrosis successfully developed by Peregrine, which has clear anti-tumor effect. However, as a mouse/human chimeric mab, CHTNT must have its own shortcomings. Humanization of murine-derived antibodies is a complicated technique, and there is still a certain degree of host antibody reaction. With the development of biotechnology, monoclonal antibody must go through the development stage from murine monoclonal antibody, mouse/human chimeric monoclonal antibody, humanized monoclonal antibody to whole human monoclonal antibody. Therefore, as a clinical therapeutic antibody, compared with chimeric antibody and humanized antibody, fully humanized antibody is undoubtedly the best choice. At present, fully human antibody preparation techniques based on antibody library technology and human-human hybridoma technology have been relatively mature, so it is necessary to prepare fully human monoclonal antibodies targeting denatured necrotic nuclei in tumor necrotic areas that can be used for TNT.

Fab fragment, also known as antigen-binding fragment, refers to the antigen-binding region of an antibody, consisting of a complete light chain and a heavy chain variable region and constant region CH1 domain. The Fab fragment contains 440~450 amino acids and its molecular weight is about 55kDa. Compared with complete IgG, Fab antibody fragments lack the constant Fc region of glycoylation, and have the advantages of strong tissue penetration, short half-life, low immunogenicity and low production cost, showing broad application prospects in the fields of diagnosis and clinical therapy. It has advantages in tumor treatment and is suitable for TNT treatment.

Alpha Lifetech Can Provide

The construction of Fab antibody libraries is one of the effective ways to obtain high quality and high affinity drug antibodies. The high library capacity of phage display Fab antibody libraries improves the speed of antibody engineering, such as the preparation speed of recombinant human monoclonal antibodies and chimeric antibodies.

The core of the phage display Fab antibody library is to increase the number of independent phage display clones. In the laboratory of Alpha Lifetech, our scientists strive to form a large number of independent clones of the antibody variable region (VH and VL) genes, especially the CDR region (complementary determination region), using different strategies and methods, such as amino acid mutation, frameshift reading frame, etc. In addition, we designed a scheme: after sensitizing human peripheral blood mononuclear cells in vitro, mononuclear cells with high affinity were isolated by flow sorting technology, and monoclonal treatment was carried out by EBV. By using this method, highly consistent mononuclear cell clones could be obtained, and then library construction was carried out.

Alpha Lifetech has extensive experience in the phage display antibody discovery. Our phage display platform is well-established and we have been able to use it for various types of phages. The M13 phage platform is the most widely used for phage display. It is also extensively used in many research areas. There are a variety of phage display services available via T4 and T7 phage system, which may offer different promising options depending on their particular merits.

Fab Antibody Humanization Service

Alpha Lifetech not only provides efficient antibody humanization services, but also has many years of practical experience in the field of recombinant antibody library construction, expression and purification. Our service is based on replacing the framework region with human sequences while maintaining the integrity of the antigen binding site, thus reducing the immunogenicity of the antibody. Our approach provides a balance between maintaining antigen specificity and reducing the risk of an immune response, ensuring a more effective and safer therapeutic outcome compared to other humanization techniques. Alpha Lifetech will provide services such as antibody sequence prediction and antibody characterization analysis throughout the entire process to ensure that our clients' projects run smoothly.

Fab Antibody Humanization Service Workflow

Steps	QC Standard	Timeline	Delivery standards
Step1: Antibody Isolation	Purity >95%	1-2 weeks	Purified Antibody
Step2: Sequence Analysis	Accurate identification of non-human sequences	1 week	Sequence Report
Step3: CDR transplantation and frame engineering	Successful integration and expression	2-3 weeks	Humanized Antibody Construction
Step4: Expression and Purification	High yield and purity	2-3 weeks	Purified Humanized Fab Antibody
Step5: Functional Validation	Affinity and specificity assessment	1 week	Validation Report