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Human scFv Antibody Library Construction Service

KMD Bioscience has been working on antibody research for many years. We have extensive experience in antibody library construction, with our well-established antibody phage display platform, KMD Bioscience can provide high-quality human scFv antibody library construction service. A single-chain fragment variable (scFv) is a recombinant antibody, because of its small molecular weight, strong tissue penetration, single-chain fragment variable (scFv) antibody has great potential for use in many fields such as research, diagnostic and clinical applications. By using advanced phage display technology, all scFv antibodies we produced guarantee high affinity and stable structure.

 

Human scFv Antibody Libraries:

 

Phage display antibody libraries are widely used in screening high affinity antibodies, such as screening of angiogenesis markers fibronectin, tumor-specific B3/B4 antigen and EGFR. Antibody libraries constructed by phage display techniques are usually derived from RT-PCR amplification of peripheral blood mononuclear cells, spleen cells, bone marrow cells and tonsil cells mRNAs to form cDNA fragments. These gene fragments were spliced into phagemid vectors, and then transferred into phages to form covalent structural protein and display in vitro. The heavy and light chains of the antibody are expressed independently and then randomly combined to form a large antibody library with phage clone numbers of 1011-1012.

 

 

The scFv antibody library refers to the variable region (VH and VL), especially the CDR region genes of the antibody, forms a large number of different clones by different strategies, such as amino acid mutations, frameshift mutations. And high affinity single chain Fv antibodies against the target antigen were screened from these clones. Since the scFv antibody has a smaller molecular weight (25 kDa), the scFV antibody is more stable and does not easily form a multimer compared with the Fab antibody library.

 

Types of Phage Display System:

 

KMD Bioscience has extensive experience in phage display platform by employing various types of phage in various project objectives. In general, M13 phage system is the most popular vehicle choice for phage display and extensively used in many types of research. We also provide phage display related services via either T4 or T7 phage systems, which can provide different promising alternatives by their special merits. Meanwhile, clients can also provide antigens, then higher affinity and specificity of monoclonal antibodies can be obtained by sensitizing peripheral blood lymphocytes in vitro.

 

Service Process:

 

 

Service Procedure:

 

Steps

Specification

Timeline

Deliverables

cDNA synthesis

1. Total RNA extraction.
2. RT-PCR.

3. PCR amplification with Actin specific primers to identify the quality of cDNA.

8-9 weeks

* Experimental report: including detailed construction procedures and representative sequence information.

 

* Product delivery: including bacterial and phage antibody libraries.

Library construction

1. Primer design & synthesis

2. PCR amplification of variable region genes of heavy and light chains using cDNA as a template.

3. scFv gene splicing.

4. Plasmid construction & transformation: after enzyme digestion, scFv and phagemid vector were ligated and transformed into TG1 host bacteria by electric shock to construct antibody libraries.

5. Identification: 20-50 clones were randomly selected, PCR identification, sequencing and analysis of antibody sequences.

6. Packaging: >1*1013 phage particles /ml.

 

Service Highlights:

 

--Extensive experience in antibody engineering development

--Guaranteed high affinity: 10-7 - 10-13M

--Multiple phage display systems: M13, T4, T7

--Large size of libraries: 107 - 1012 independent clones

--Strict quality assurance: cGMP management rules

--Individuation library searching strategies

--One-stop services, including antibody development, antibody sequencing, antibody labeling and related services