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Phage Display Antibody Development Platform

Based on the comprehensive platform system construction of antibody platform,  Alpha Lifetech can provide technical services from antibody preparation, antibody purification, antibody sequencing, etc.

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Phage Display Antibody Development Platform


Alpha Lifetech, with its extensive project experience in antibody discovery, can provide customized monoclonal and polyclonal antibody services for multiple species, as well as antibody library construction and screening services. Alpha Lifetech can provide customers with quality assured antibody and recombinant antibody products, as well as corresponding services, to prepare efficient, highly specific, and stable antibodies. By utilizing comprehensive antibody platforms and antibody engineering, we provide services covering the upstream and downstream of antibody production, including technical services such as antibody preparation, purification, antibody sequencing, and validation.

Alpha Lifetech has a mature antibody discovery platform that provides customers with a range of services including animal immunity, phage display antibody library construction and screening, antibody sequencing, antibody expression, antibody purification, antibody validation, and antibody labeling based on hybridoma technology, single B cell technology, phage display technology, and more. Alpha Lifetech has a standard production line, and in addition to antibody preparation services, supporting services such as antibody humanization, antibody affinity maturation, ADC drug design and development, and CAR-T subsequent sequence design can also be provided. At the same time, Alpha Lifetech has constructed an antibody library based on M13, T4, and T7 phage display technology platforms, with a storage capacity of up to 10^8-10^9. The positive rate, insertion rate, and diversity of the library can all reach over 90%.

Phage display Technology

Monoclonal antibodies production was initially prepared using hybridoma monoclonal antibody technology to produce mouse hybridoma monoclonal antibody. Specifically, by fusing spleen cells from immunized mice with myeloma cells from humans or mice, hybridoma cells are formed, which secrete specific hybridoma monoclonal antibody antibodies. By humanized modification of mouse monoclonal antibodies and genetic engineering modification of the antibodies to give them a constant human immunoglobulin region, their immunogenicity can be reduced. Monoclonal antibody engineering are widely used in biomedical and clinical applications.

Alpha Lifetech can provide one-stop antibody engineering services from preclinical research protocol design to antibody drug conjugates (ADCs) discovery and animal validation. Antibody drug conjugates (ADCs) can deliver chemotherapy drugs to cancer cells. After binding to specific targets expressed on cancer cells, ADC releases cytotoxic drugs into cancer cells. Alpha Lifetech can also provide customers with comprehensive antibody affinity maturation services. With advanced mutation optimization and high-throughput phage display screening technology, antibodies with certain affinity are first screened, and then amino acid mutations are introduced to generate diverse variants. Subsequently, high affinity antibodies were evaluated and screened using screening techniques. After multiple rounds of optimization and structural analysis, antibodies with higher affinity and stronger specificity were ultimately obtained.

Alpha Lifetech can construct various types of antibody phage display libraries, including immune libraries, native libraries, semi synthetic libraries, and synthetic libraries. Based on the high capacity of antibody libraries, highly specific antibodies can be obtained. Multiple phagemid vectors such as pMECS, pComb3X, and pCANTAB 5E can be provided, as well as strains such as TG1 E. coli, XL1 Blue, and ER2738. In addition to a library capacity of up to 10^9, the target fragment insertion rate of the library is also high, creating sufficient conditions for screening antibodies that satisfy customers. The flowchart of Alpha Lifetech's antibody preparation service based on phage display technology is shown in Figure 1.

phage display-Alpha Lifetech
Fig.1 Process diagram for preparing monoclonal antibodies based on phage display technology. 

Phage Display Antibody Production Workflow

Steps Service Content Timeline
Step 1: Animal immunization
(1) Animal immunization 4 times, booster immunization 1 dose, a total of 5 doses immunized.
(2) Negative serum before immunization was collected, and ELISA was performed on the fourth dose to detect serum titer.
(3) If the serum antibody titer of the fourth dose meets the requirements, one additional dose of immunization will be administered 7 days before blood collection. If it does not meet the requirements, routine immunization will continue.
(4) Qualified potency, blood collection and separation of monocytes
10 weeks
Step 2: cDNA Preparation
(1) PBMC Total RNA Extraction (RNA Extraction Kit)
(2) High fidelity RT-PCR preparation of cDNA (reverse transcription kit)
1 day
Step 3:  Construction of Antibody Library
(1) Using cDNA as a template, genes were amplified by two rounds of PCR.
(2) Phage construction and transformation:  gene splicing phagemid vector, electroporation transformation of TG1 host bacteria, construction of antibody library.
(3) Identification: Randomly select 24 clones, PCR identification positive rate+insertion rate.
(4) Assisted phage preparation: M13 phage amplification+purification.
(5) Phage display library rescue
3-4 weeks
Step 4: Antibody Library Screening (3 rounds)
(1) Default 3-round screening (solid-phase screening): pressure screening to remove non-specific antibodies to the maximum extent possible.
(2) Single clone amplification bacteriophage selected+IPTG induced expression+ELISA detection of positive clones.
(3) All positive clones were selected for gene sequencing.
4-5 weeks

Advantage Of phage Display Technology

Alpha Lifetech has rich experience in the field of antibody development. Over the years, Alpha Lifetech has established a comprehensive antibody development platform.

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Support Services

We can provide different animal based immune library construction services and natural antibody library screening services according to customer needs

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Multiple Target

Multiple target antibody discovery services are available: proteins, peptides, small molecules, viruses, membrane proteins, mRNA, etc.

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Multiple Vectors

Personalized library construction service, we can provide various bacteriophage vectors including PMECS, pComb3X, and pCANTAB 5E, and modify them according to customer needs

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Mature Platform

Storage capacity can reach 10 ^ 8-10 ^ 9, insertion rates are all above 90%, and the affinity of antibodies obtained through screening is generally at the nM pM level

Related SERVICE

Multiple Antibody Development Strategies

monoclonal antibody development-Alpha Lifetech

Monoclonal Antibody Development Service

We can provide high-quality, high-purity, and highly specific monoclonal antibody development services, including the production of mouse monoclonal antibodies and rabbit monoclonal antibodies

hybridoma cells-Alpha Lifetech

Hybridoma Technology Platform

Including immunization program, antibody preparation services, antibody purification, antibody high throughput sequencing, antibody validation, etc

b cell screening-Alpha Lifetech

Single B Cell Sorting Platform

Alpha Lifetech has advantages in screening time and obtaining high-quality antibodies. It can provide antigen design, synthesis, and modification, animal immunity, single B cell enrichment screening, single cell sequencing.

phage display-Alpha Lifetech

Phage Display Antibody Development Platform

Alpha Lifetech can provide phage display antibody development technical services from antibody preparation, antibody purification, antibody sequencing, etc.

If you have any questions, please feel free to contact us at any time.

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