AtaGenix Laboratories

Immune or Naive Phage Display Libraries available for screening
Can develop Monoclonal Antibodies from our Phage Display Libraries

Description

Stable transfection is the process of integrating exogenous plasmid DNAs into chromosomes of host cells, causing the host cells to express target gene and protein for long periods of time. The target cells will be screened based on transient transfection. To build stable cell lines, target cells are screened by related drugs according to the resistance marker contained in the vectors. The most commonly used eukaryotic expression vector gene markers are hygromycin, neomycin, puromycin and Hygromycin B. G418 and puromycin are commonly used for selective screening. The selected cell lines can stably express the target protein.

Provisions by AtaGenix
AtaGenix can either construct vectors or provide stable selection of the built foreign plasmid DNA. Corresponding drugs will be selected according to the resistance markers of the plasmid DNA to carry out drug test on the cells to be screened and select the lowest concentration when all the cells undergo apoptosis. Upon transient transfection, the infected cells will be screened by a suitable concentration of drugs. After 10-14 days, single clones will be picked out for amplification to detect protein expression by ELISA, SDS-PAGE and Western Blot etc. Finally, stable cell lines can be screened.

Process Flow of Stable Cell Line Service

1. Vector construction: Exogenous genes are inserted into appropriate vectors.
2. Determination of screening concentration: The concentration of antibiotics for all the cells which died over a 10-14-day period is determined as the screening concentration.
3. Cell inoculation: Cell inoculation should be carried out two days earlier than the transfection operation. Various cell densities on the plates are dependent on their growth rates and shape of the cells. Cells should cover 60% - 80% of the plate on the day of transfection.
4. Cell transfection: Transfection of target cells is handled with built vectors.
5. Antibiotic screening.
6. Identification of screening results.

Submission by Customers

1. Constructed vectors containing extraneous gene or gene templates.
2. Information about cell lines to be built.

AtaGenix's Advantages

1. Stability: We guarantee stable expression of transfected cells within 15 generations.
2. Economical: Affordable prices, high cost performance.
3. Quality assurance: Strict identification of exogenous genes.

Deliverables

1. Vectors containing exogenous genes
2. Stable cell lines.

AtaGenix provides two methods for large-scale production of monoclonal antibodies, ascites preparation and roller bottle culture.

AtaGenix's Advantages
Professional SPF animal laboratory, with sterile roller bottle culture apparatus in large scale．
Ascites can be rapidly and efficiently obtained for hybridoma cultivation．
Sophisticated technicians will handle with the purification of ascites and supernatant of hybridoma culture.

Mouse or Rabbit

Deliverables:

1. 1-2mg ProteinA/G purified monoclonal polyclonal by purification, and the purity is above 90% (If the client needs the antigen purified antibody, AtaGenix is able to provide such a service as required).
2. 1-2 hybridoma cell strains
3. ELISA efficacy of purified antibody is above 1:64000, and we guarantee the WB test is antigen positive.
4. Test report, which contains SDS-PAGE and QC of the final concentration

AtaGenix's Advantages

1. Advanced antibody production techniques conforming to international standards
2. Premium and varied antigens
   (A) Recombinant protein
   (B) Peptide
   (C) Small molecules
   (D) Others
3. Service items are diversified and charged according to the project stage, which could save cost for client.
4. Each project will be overseen by one specific person, including the project negotiations, progress of the project and the project deliverables. The client will then be given the opportunity to test the final product, resulting in high customer satisfaction.
5. AtaGenix offers a high-quality antibody recombinant expression service.

Requirement of the samples provided by clients:

1. Client provides the purified antigen, and the purity should be above 60% and the amount reaches 3mg (2.5mg protein for immunization and 0.5mg protein for screening)
2. Client provides the expression plasmid, information about the plasmid as well as the clone scheme.

AtaGenix offers a one-stop service for custom antibody preparation, from antigen preparation, to antibody production, antibody labelling service as well as antibody sequencing service.

AtaGenix is able to provide clients with a tailored polyclonal antibodies service in different hosts, including rabbit, mouse, rat, guinea pig, goat and chicken.

Polyclonal Antibodies Preparation contains:
√　Antigen preparation
　　(1) Peptide antigen synthesis
　　(2) Protein antigen preparation
　　(3) Other antigens
√　Immunization - AtaGenix will provide an optimised and tested immunization scheme, or complete the immunisation process according to the requirements of the client
√　Antiserum production
√　Antibody purification
　　(1) Protein A/G purification
　　(2) Affinity purification