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Email:info@kmdbioscience.com

Phosphorylated Antibody Production Service

KMD Bioscience offers first-in-class custom antibody services. Such as phosphorylation-specific polyclonal and monoclonal antibody services with a success rate of 100%. That includes synthesis of phosphorylated and non-phosphorylated peptides, conjugation to a carrier protein, immunization, serum production and affinity purification of the antibodies.

Phospho-specific Antibody Production

Protein phosphorylation is a very important post-translational modification (PTM) in cells. It mainly involves the regulation of the protein biological activity, including activation or inhibition of protein activity and ultimately regulating the cell signal transduction pathway. KMD Bioscience can provides phosphorylation-specific antibody production services based on phage display platform.

 

Service Procedure:

Steps

Specification

Deliverables

Antigen design and synthesis

* Clients provide target antigen (polypeptide, protein), purity >90%.
* We designed and synthesized phosphorylated or non-phosphorylated antigen peptides. (Optional)

(Antigen production by KMD Bioscience)

* MS and HPLC reports (for both phosphopeptide and non-phosphopeptide).

* 2-5mg phosphopeptide (>90% purity).

* 2-5mg non-phosphopeptide (>90% purity).

Animal immunization

* Protocol 1: Immunization of 5 Balb/c mouse (using a unique mouse immunoadjuvant to produce high titer antiserum rapidly in 20 days). Detection of serum titer by ELISA.
* Pr
otocol 2: Immunization of 2-3 rabbits (using a unique rabbit immunoadjuvant to produce high titer antiserum rapidly in 28 days). Detection of serum titer by ELISA. (Other animals are available)

* 1-2 ml of pre-immune serum.
* ELISA
identification report.

Production of antibody phage display libraries

1. Collecting B lymphocytes after immunization.

2. mRNA extraction & cDNA synthesis & phage vector construction & transformation & amplification.

3. Screening of antibody library with phosphorylated and non-phosphorylated antigen.

4. Amplification of positive clones & Construction of shuttle plasmids.

5. Small-scale and large-scale mammalian system expression (293, CHO). Antibody purification and identification.

* Recombinant expression plasmid.
* 2-5 mg of purified recombinant antibody.

* ELISA results (both for phosphopeptide and nonphosphopeptide).

* Western Blot identification report (Optional).

 
Case 1:

Service Highlights:

*Extensive experience: We have many years’ experience and a strong track record in antibody production and phage display platform.

*High specificity: The recombinant monoclonal antibody only recognizes specific phosphorylation sites and does not react with non-phosphorylated antigen.

*GMP/GLP grade.

*ELISA titer up to ~105.

*A variety of production modes: 5L, 15L, 80L fermenters for selection.

 

In addition, we also provide phosphorylation-specific polyclonal and monoclonal antibody services based on traditional hybridoma and polyclonal antibody production technologies.