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Phosphorylated antibody preparation service
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Phosphorylated antibody preparation serviceIntroduction

Leveraging years of R&D expertise and extensive project experience, KMD Bioscience's antibody development platform has established mature capabilities in generating phosphorylation-specific antibodies. The phosphorylation-specific polyclonal and monoclonal antibodies we develop demonstrate performance comparable to that of comparable products from Cell Signaling Technology (CST). We offer clients a one-stop, end-to-end service for phosphorylation antibody preparation, covering phosphorylated peptide antigen design, customized experimental protocol planning, and multi-dimensional antibody validation. Our validation system includes cross‑reactivity testing against phosphorylated versus non‑phosphorylated antigens, as well as application assessments in Western blotting and immunohistochemistry, thereby fully addressing the diverse project requirements of different clients.

Protein phosphorylation is a core class of post‑translational modifications (PTMs) within cells. By mediating the activation or inhibition of target protein biological activities, it precisely regulates the operation of intracellular signal transduction pathways and represents a critical molecular mechanism governing cellular functions. In eukaryotes, protein phosphorylation predominantly occurs on serine, threonine, and tyrosine residues; histidine phosphorylation, in contrast, is the most representative form of phosphorylation regulation in prokaryotes. Leveraging our well-established phage display technology platform, KMD Bioscience further provides customized preparation services for phosphorylation‑specific recombinant monoclonal antibodies to meet specialized project needs.


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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