Antigen Design & Synthesis

* Client provides antigen (peptide/protein) with >90% purity
* KMDTECH designs and synthesizes phosphorylated/non-phosphorylated antigens (optional)

(If prepared by KMDTECH)
* MS and HPLC reports for phosphorylated/non-phosphorylated antigens
* 5 mg phosphopeptide (>90% purity)
* 5 mg non-phosphorylated peptide (>90% purity)

Antigen Conjugation

* Client provides conjugation antigens (5 mg KLH/OVA-conjugated phosphopeptide, 5 mg BSA-conjugated phosphopeptide, 5 mg BSA-conjugated non-phosphorylated peptide)
* KMDTECH performs peptide conjugation (optional)

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

Animal Immunization

Option 1: Immunize 5 Balb/c mice (using mouse-specific adjuvant for rapid high-titer serum generation within 20 days), ELISA serum titer detection
Option 2: Immunize 2-3 New Zealand White rabbits (using rabbit-specific adjuvant for high-titer serum generation within 28 days), ELISA serum titer detection (other species optional)

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

Phage Display Antibody Library Preparation

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

* Recombinant expression plasmid
* 2-5 mg purified recombinant antibody
* ELISA report (including titer and cross-reactivity report)
* Antibody titer ≥10⁴ against phospho-antigen with no reaction to non-phosphorylated antigen (ELISA)
* Western Blot validation report (optional)