Phage Display Service

M13 single strand filamentous phage display system is the most widely used library display system. The phage display library is formed by fusing the protein, antibody or polypeptide with phage pIII protein to participate in phage package and display on the phage surface. Using phage display library to make 3-5 rounds of affinity screening for specific target molecules, phage that specifically binds to the target molecule could be enriched to obtain corresponding DNA sequence information.

Our current phage display technology service platform covers frequently-used library types such as immune antibody library and natural antibody library. We strive for fulfilling individual customer's needs by customizing a variety of library and screening services to fit the individual preference with flexibility and efficiency.

Phage Display Service Profile

Leading Biology platform system mainly includes pComb3 phage display system and pCANTAB5e phage display system.

pComb3 Phage Display Technology

(1) Obtain the gene library: Collect the immune cells from the target antibody host and extract the total RNA of the cells. Amplify VH-CH1 antibody gene fragment and VL-CL1 antibody gene fragment respectively with multiple pairs of primers after reverse transcription.

(2) Transform E.coli with antibody gene library: The amplified antibody gene library was ligated to pComb3 plasmid, and then electrotransform E.coli XL12 Blue. Identify the capacity of gene library and the insert rate of antibody gene.

(3) Phage Fab antibody library construction: The helper phage VCSM13 was added to E.coli which was transferred into antibody gene and grown to logarithmic phase in order to rescue the phage. After overnight culture, collect the supernatant by centrifugation and get the phage Fab antibody library.

(4) Antibody library titer determination: Gradient dilute the phage Fab library obtained above to perform plaque culture experiment. Observe the growth of plaque to determine the titer of phage Fab antibody library.

(5) Enrichment screening of phage antibody library: Add phage antibody to the target antigen coated enzyme-linked plate to incubate, then follow by washing and eluting. Neutralize the eluted phage antibody, infect the bacteria again and carry out the second round of elutriation. Three rounds of elutriation would be conducted and titer was measured after each round of elution. During the last screening, set a BSA-coated control group to determine whether a large amount of anti-BSA phage antibody was enriched.

Fig 1. Process of pComb3 phage Display Technology

Fig 2. Genetic Map of Phagemid Vector pComb3

Fig 3. Genetic Map of Helper Phage VCSM13

pCANTAB5e Phage Display Technology

(1) Obtain the ScFv gene library: Collect the immune cells from the target antibody host and extract the total RNA of the cells. Amplify VH antibody gene fragment and VL antibody gene fragment respectively with multiple pairs of primers after reverse transcription. Randomly splice the full set of VH and VL gene into ScFv gene library with Linker primers after VH and VL fragment purification and recycling.

(2) Transform E.coli with ScFv gene library: The full set ScFv gene fragment was ligated to pCANTAB5e vector, and then electrotransform E.coli TG1. Identify the capacity of ScFv gene library and the insert rate of gene.

(3) Phage ScFv antibody library construction: The helper phage M13K07 was added to E.coli which was transferred into ScFv antibody gene and grown to logarithmic phase in order to rescue the phage. After overnight culture, collect the supernatant by centrifugation and get the phage ScFv antibody library.

(4) Antibody library titer determination: Gradient dilute the phage ScFv antibody library obtained above to perform plaque culture experiment. Observe the growth of plaque to determine the titer of phage ScFv antibody library.

(5) Enrichment screening of phage antibody library: Add phage antibody to the target antigen coated enzyme-linked plate to incubate, then follow by washing and eluting. Neutralize the eluted phage antibody, infect the bacteria again and carry out the second round of elutriation. Three rounds of elutriation would be conducted and titer was measured after each round of elution. During the last screening, set a BSA-coated control group to determine whether a large amount of anti-BSA phage antibody was enriched.

Fig 4. Process of pCANTAB5e phage Display Technology

Fig 5. Genetic Map of Phagemid Vector pCANTAB5e

Fig 6. Genetic Map of Helper Phage M13K07

Phage Display Technology Service Items

(Phage Display Antibody Library Construction)

Phage Display Library Screening

Our current phage display technology service platform covers commonly used library types such as immune antibody library and natural antibody library. We will provide flexible and efficient screening services according to the self-own phage library products chosen by clients or various personalized library provided by Leading Biology.

 

The following experiment procedures would be carried out according to the samples provided by clients:

(1) Antibody library titer determination: Gradient dilute the phage ScFv antibody library obtained above to perform plaque culture experiment. Observe the growth of plaque to determine the titer of phage ScFv antibody library.

(2) Enrichment screening of phage antibody library: Add phage antibody to the target antigen coated enzyme-linked plate to incubate, then follow by washing and eluting. Neutralize the eluted phage antibody, infect the bacteria again and carry out the second round of elutriation. Three rounds of elutriation would be conducted and titer was measured after each round of elution. During the last screening, set a BSA-coated control group to determine whether a large amount of anti-BSA phage antibody was enriched.

Table 2. Phage Display Technology Service Items: Antibody Screening

Related Services

Premade Phage Display Antibody Libraries

Phage Display Library Construction

Phage Display & Antibody Library Services

Why Leading Biology?

With over 5 years of experience in custom antibodies, we offer numerous benefits for developing your custom antibody:

• Our industry leading titer guarantees of 1:50,000 eliminate the risk of not obtaining antibodies against peptide antigens.

• Our economies of scale allow us to pass cost savings to you and help maximize your budget.

• 100% of services are transparent throughout.

• Antibodies from our facility have been cited in many published research papers.

• By outsourcing antibody production and characterization needs to us, you can spend more time focused on your research.

• Our ideal location in California ensures that animals benefit from outdoor facilities and mild year-round temperatures.

• Confidentiality is a top priority for all projects. Antibodies that we manufacture belong to you and will not be commercialized.

Contact Information

Please obtain a quote before ordering, and refer to the quote number when you place an order.

Orders are typically confirmed within 12 hours.

Have a Question? Email us info@leadingbiology.com

Order Products: Order Related Products

By Phone: 1-661-524(LBI)-0262 (USA)