Phage display technology was first established in 1985 and was used 5 years later for discovering antibodies. It is now considered as a revolutionary breakthrough. Phage display antibody technology helps researcher to acquire antibody from animals that does not have a stable myeloma cell line thus enlarging the potential types of host animal. Another significant improvement is the ability to keep all of the antibody sequences from immune-responded cells for downstream screening. A naïve antibody library is an antibody library amplified from a natural source, such as primary B-cells of non immunized donors, through phage display. Bio Bench 100-HuAb Human naïve antibody library is successfully built up with PBMCs from 3000 healthy human donors.
Workflow of Phage Display Antibody Library
Considering animal life respecting purpose, immunogenicity issue and/or toxicity reaction, to find the affine antibody from the naïve antibody library is a greater approach, compare to hybridoma antibody technology. Since big data analysis and computer modeling technology supporting biotech research, to build up antibody library based on data become possible. A synthetic antibody library is based on computational in silico design and gene synthesis and the CDR design and composition is precisely defined and controlled. Semi-synthetic libraries comprise both CDRs from natural sources as well as in silico design of defined parts.
Typical process for your phage display antibody project (the exact procedure may be different)
Phase 1 Antigen
Gene cloning (if needed).
Mammalian cell expression.
1.QC Report. 2.100μg peptide or 1mg protein. 3.Cell line (if customer requires).
Peptide 1 week. Protein expression 7 – 12 weeks.
Phase 2 Immunization
DNA/Peptide/protein/whole cell immunization.
Immunize 3, 5, 7 or 10 BALB/c mouse.
Immunize 2, 3, or 5 New Zealand rabbit.
Immunize 1, 2, or 3 llama.
Bio Bench enhanced adjuvant.
1.Test bleed report, titer 1:64 000. 2.10μL serum sample at final boost or earlier (if customer requires).
4 – 8 weeks.
Phase 3 Antibody library construction
Cell isolation (spleen, bone marrow, PBMC).
VH and VL PCR amplification.
Phagemid synthesis and cloning.
1.Parental supernatant test report. 2.2 mL supernatant (if customer requires).
8 – 13 weeks.
Phase 4 Biopanning
2 - 6 rounds of biopanning through protein, cell based and/or in vivo screening.
Screening and validation by high throughput platform (ELISA, FACS, etc.) until at least 10 different binders are identified.
1.Test report. 2. 5mL supernatant of the subclones. 3. 2 vials for each clone. 4. 1mg/clone purified antibody.
3 – 6 weeks.
Phase 5 Antibody DNA sequencing
Phage DNA extraction and antibody sequencing.
1. Test report.
2 – 8 weeks.
Phase 6 Screening
Additional ELISA or sandwich ELISA.
Cell-based ELISA or FACS.
Other test by demand.
To be determined.
Phage display is significantly more challenge to achieve, compare to hybridoma method. It requires scientific knowledge, experienced team member, technology management and know-how solutions. Bio Bench has successfully build up several naïve antibody libraries and synthetic/semi-synthetic antibody libraries from human, llama (Alpaca) and rabbit. More than 50 drug discovery projects have been accomplished through our technology platform. Find out case studies? Please roll down the page to download them.
Immunized Antibody Library
Naive Antibody Library
Synthetic Antibody Library
Service and Time Line
AB210102 100-million clones mouse antibody library construction and screening
100-million clones ELISA KD<20nM
Peptide/protein. DNA/peptide/protein sequence. Stable cell line. NOTE: Please contact us for a detailed “list of material” regarding to your unique project.
•Acquired antibody gene sequences. •100μg purified antibody for each clone. •QC and Report.