Technical Service

Human Recombinant scFv Phage Display Antibody

Development

Antibodies are important effector molecules of the body's immune system, which can specifically recognize antigens and play an important role in modern medicine, and are widely used in basic research. Antibody development has gone through four stages: hybridoma monoclonal antibody, chimeric antibody and humanized modified monoclonal antibody, homogeneous recombinant antibody technology, and natural homogeneous recombinant antibody. Phage display antibody library technology offers the possibility of preparing homogeneous human recombinant monoclonal antibodies, which will be more and more emphasized in the development and production of antibody drugs in the future.

AtaGenix offers panning technology service for fully human recombinant scFv phage antibody library.

Advantages

1. High sequence accuracy and capacity (1011pfu) .

2. Two-step construction protocol to avoid mismatches introduced by PCR. The light chain library is built in advance and then heavy chains are cloned by enzymatic reactions.

3. Different from most commercially available libraries, the single-chain antibody library (scFv) is built into two light chain subtypes, κ and λ. Both libraries can be used separately.

4. All sequences are HIS tagged,expressed directly by using phagemid and purified by affinity chromatography.

QC Testing

1. Total Insertion Rate and Accuracy

The fragment insertion rate is 100%, and the antibody sequence accuracy is higher than 90%.

2. Antibody Sequence Diversity Analysis

2.1Germline Gene Distribution Characteristics

The fully human recombinant antibody library is divided into two light chain subtypes, κ and λ. The distribution ratio of light chain germline genes of the two subtypes and the distribution ratio of heavy chain germline genes are analyzed, indicating that the library germline gene coverage is high. The distribution characteristics are in line with the germline gene distribution law of antibody drugs.



K-type Light Chain Germline Gene Distribution Ratio



λ-type Light Chain Germline Gene Distribution Ratio


Heavy Chain Germline Gene Distribution Ratio


2.2 CDR3 Length Distribution Analysis

Sequencing analysis is performed by randomly selecting 500 clones from each of the K and λ subtype libraries with no repeats. Its CDR3 length distribution conforms to the normal distribution, which is consistent with the CDR3 length distribution law of the fully human recombinant antibody drug.



K-type Light Chain Antibody CDR3 Length Distribution Ratio

Length of VL-CDR3

  


λ -type Light Chain CDR3 Length Distribution Ratio

  


Heavy Chain CDR3 Length Distribution Ratio

Contents

Receivables

Contents

Time Frame

Deliverables

Antigen

Protein, Peptide, Small Molecule

3-4 Rounds of Panning

Polyclonal ELISA Validation

Monoclonal ELISA Validation

Positive Clone ELISA Validation

Sequencing

Experimental Report

   2-3 Weeks

   1-3 Positive Strains

   1-3 scFV Sequences

Service Report

Optional services: Antigen preparation/full length antibody production/affinity sequencing/stable cell line

construction/bispecific antibody development

Case Study


Results of Full Human Recombinant Phage Library Screening by AtaGenix