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Human scFv Antibody Library Construction Service

KMD Bioscience has a mature antibody production platform with advanced antibody engineering technology and extensive experience in antibody library construction. Based on these, KMD Bioscience can provide high-quality human scFv antibody library construction services and give you reliable data feedback. Single chain antibody fragment (scFv) is a recombinant antibody, with the advantages of small molecular weight, high tissue penetration and easy genetic engineering, it 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 marker fibronectin, tumor-specific B3/B4 antigen and EGFR. Antibody libraries constructed by phage display technology are usually derived from RT-PCR amplification of mRNAs from peripheral blood monocytes, spleen cells, bone marrow cells and tonsils to form cDNA fragments. These gene fragments were spliced into phagemid vectors, and then transferred into phages to form covalent structural protein expression and in vitro display. The heavy and light chains of the antibodies are expressed independently and then randomly combined to form a large antibody library with phage clone numbers up to 1011-1012.

 

 

The scFv antibody library refers to the variable region (VH and VL), especially the CDR region genes of the antibody, which forms a large number of different clones by different strategies, such as amino acid mutations and frameshift mutations. And high- affinity single-chain Fv antibodies against the target antigen were screened from these clones. Compared to the Fab antibody library, scFv antibodies are more stable and less prone to form polymers due to their smaller molecular weight of about 25kDa.

 

Types of Phage Display System:

 

KMD Bioscience has a well-established phage display platform and has used various types of phages for different project objectives. In general, the M13 phage system is the most popular choice for phage display and is extensively used in many types of research. KMD Bioscience also provides phage display -related services via either T4 or T7 phage systems, which can provide different promising alternatives by their special merits. KMD Bioscience supports customer-tailored services. You can provide antigens, and we perform in vitro peripheral blood lymphocyte mononuclear cell sensitization to obtain higher affinity and specificity monoclonal antibodies.

 

Service Process:

 

 

Service Procedure:

 

Steps

Specification

Timeline

Deliverables

cDNA synthesis

* Total RNA extraction.
* RT-PCR.

* 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

* Primer design & synthesis

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

* scFv gene splicing.

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

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

* 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

 

How to Order?

If you have any questions regarding our services or products, please feel free to contact us by E-mail: info@kmdbioscience.com or Tel: +86-400-621-6806