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Phosphorylated antibody preparation service
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Phosphorylated antibody preparation serviceIntroduction

    After years of project development accumulation, the antibody development platform of KMD Bioscience Technology has developed polyclonal and monoclonal antibodies with development performance comparable to the phosphorylation of Cell Signal Transduction Company (CST). We can provide customers with one-stop phosphorylated antibody preparation services, including phosphorylated peptide design, experimental protocol design and related validation services, phosphorylated and non phosphorylated antigen cross reactivity, Western Blot and immunohistochemistry, etc., to meet the project needs of different customers.

    Protein phosphorylation is an important post-translational modification (PTM) in cells, mainly involving the regulation of protein biological activity levels, including activation or inhibition of protein activity, thereby further regulating cellular signaling pathways. In eukaryotes, protein phosphorylation sites are serine, threonine, and tyrosine; The main phosphorylation site in prokaryotes is histidine. KMD Bioscience can provide customers with services for the preparation of phosphorylated monoclonal recombinant antibodies based on phage display technology.

Service content and process

Step

Service Content

Deliverables

Antigen Design & Synthesis

* Customer provides antigen (peptide, protein), purity >90%
* KMD Biosystems designs and synthesizes phosphorylated and non-phosphorylated antigens (optional)
(Antigen prepared by KMD Bioscience)

* MS and HPLC reports for phosphorylated and non-phosphorylated antigens
* 5 mg phosphorylated peptide, purity >90%
* 5 mg non-phosphorylated peptide, purity >90%

Antigen Conjugation

* Customer provides conjugated antigen (KLH/OVA-conjugated phosphorylated peptide, 5 mg; BSA-conjugated phosphorylated peptide, 5 mg; BSA-conjugated non-phosphorylated peptide, 5 mg)
* KMD Biosystems performs peptide conjugation (optional)

* 5 mg KLH/OVA-conjugated phosphorylated peptide, purity >90%
* 5 mg BSA-conjugated phosphorylated peptide, purity >90%
* 5 mg BSA-conjugated non-phosphorylated peptide, purity >90%

Animal Immunization

* Option 1: Immunize 5 Balb/c mice (using mouse adjuvant, high antibody titer serum in 20 days), ELISA for serum titer
* Option 2: Immunize 2-3 New Zealand rabbits (using rabbit-specific adjuvant, high antibody titer in 28 days), ELISA for serum titer (other animals available)

* 1-2 ml pre-immune serum
* ELISA serum titer validation report

Antibody Phage Display Library Construction

* Collect immune B lymphocytes
* Extract mRNA, PCR amplify cDNA, construct phage vector, transform, amplify
* Positive and negative screening of antibody library with phosphorylated and non-phosphorylated antigens
* Amplify positive clones, construct shuttle vector
* Small-scale and large-scale mammalian expression (293, CHO), antibody purification and validation

* Recombinant expression plasmid
* 2-5 mg purified recombinant antibody
* ELISA report including titer and cross-reactivity data
* Antibody titer against phosphorylated antigen ≥10^4 with no reactivity to non-phosphorylated antigen (ELISA)
* Western Blot validation report (optional)


Service Advantages

Advantages of Phosphorylated Antibody Preparation Services-EN.png

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