Phospho-Specific Monoclonal Antibody

Monoclonal Antibody

A type of protein made in the laboratory to bind to the substances in the body. It is used alone or to carry drugs, toxins, or radioactive substances directly to cancer cells. They can recognize and attach to specific proteins produced by cells.

Types of Monoclonal Antibody

  • Naked monoclonal antibodies ― antibodies without any drug or radioactive material attached to them. This antibody attaches to cancer cells to act as a marker for the body’s immune system to destroy them.
  • Conjugated monoclonal antibodies ― antibodies that joined to a chemotherapy drug, radioactive particle or cancer cell-killing agent. This lessens the damage to normal cells in other parts of the body caused by the drug, cancer killing agent or radioactive substances.

Phase I: Immunogen Preparation (28 days)

  • Peptide synthesis: 5 mg of phospho-peptide with >90% purity and 3 mg non-phosphopeptide with >70% purity is synthesized. 2 mg of phospho-peptide and 2 mg of non-phospho-peptide will be delivered to the customer along with the final antibody product.
  • Conjugation: The modified peptide is conjugated to the carrier protein KLH.

Phase II: Immunization and cell infusion (45 days)

  • Immunization: Balb/c mice or rats are immunized and boosted with MonoExpress immunization protocol.
  • Fusion: Electrofusion is performed at least 10,000 clones can be obtained from each fusion.

Phase III: Screening and supernatant samples preparation for 1st delivery (45 days)

  • Screening and isotyping: The hybridoma cells are screened by ELISA with phospho- and non-phosphopeptides and all positive clones are expanded into 24-well plates. Up to 5 clones with the best ELISA value and specificity to phosphopeptide are selected for isotyping.
  • 1st delivery: 2 mL supernatant samples for each clone are prepared for 1st delivery. Antibody Certificate of Analysis report is prepared with ELISA data and IgG concentration for each sample.

Phase IV: Subcloning, expansion, cryopreservation, antibody production and final delivery (30 days)

  • Subcloning, expansion, and cryopreservation: Up to 5 clones are subcloned by limiting dilution, expanded and frozen.
  • Antibody production: At least 2 mg purified antibody is produced for the clone with best screening result. The cross-reactivity with non-phosphopeptide < 10% for the purified antibody.
  • Final Delivery: Two vials of cell culture for each cell line, at least 2 mg qualified purified antibody, 2 mg of unconjugated phospho-peptide and 2 mg of a non-phosphopeptide are delivered to the customer.

Possible Effects of Monoclonal Antibody

Monoclonal antibodies are given intravenously, these antibodies are proteins thus, the human immune system may see these antibodies as foreign and it will mount a response, the possible effects include:

  • Fever
  • Chills
  • Weakness
  • Headache
  • Nausea
  • Vomiting
  • Diarrhea
  • Low blood pressure
  • Rashes