Resources » Technical Resource Centers » Antibody Technical Resources » Technology and Protocols » Monoclonal Antibody production protocol
GenScript Share

Monoclonal Antibody Development Protocol

MonoExpress™ Gold (SC1572P)

MonoExpress™ Silver (SC1661)

MonoExpress™ Bronze (SC1660)

MonoExpress™ Basic (SC1572)

MonoExpress™ Phospho-specific Antibody (SC1659)

Note:

GenScript provides free cell line storage service for 6 months from the date of cell freezing. If continued storage service is required after the end of six-month period, the charge is set at $10 per cell line per year.

MonoExpress™ Gold (SC1572P)   Fastest Custom Monoclonal Antibody Services
 
Description
Time Line
Phase I: Immunogen preparation
  • Protein expression: Full length or one protein domain is expressed in E.coli system by GenScript as immunogen according to the analysis in quotation step. 200µg immunogen protein will be delivered to the customer along with the final antibody product.
35 days
Phase II: Immunization and cell fusion with MonoExpress™ protocol
  • Immunization: Balb/c mice or rats are immunized and boosted with MonoExpress™ immunization protocol.
  • Fusion: Electrofusion is performed and at least 10,000 clones can be obtained from each fusion. Additional fusions are available to reach our guarantee without extra charge.
45 days
Phase III: Screening and supernatant samples preparation for 1st delivery
  • Screening and isotyping: Indirect ELISA is used for screening with immunogen and positive clones are expanded. 10 clones with best screening result are selected for isotyping.
  • 1st delivery: 2ml 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.
  • Cell line maintenance: The clones selected are maintained to wait for the evaluation result from the customer.
Phase IV: Subcloning, expansion, cryopreservation, antibody production and final delivery
  • Subcloning, expansion and cryopreservation: Up to 5 clones are subcloned by limiting dilution according to the evaluation result from the customer, expanded and frozen.
  • Antibody production: At least 2 mg purified antibody is produced for one clone selected by the customer.
  • Final Delivery: 200µg immunogen protein, two vials of cell culture for each cell line and at least 2 mg purified antibody are delivered to the customer.
 30 days

MonoExpress™ Silver (SC1661)   Fastest Custom Monoclonal Antibody Services
 
Description
Time Line
Phase I: Immunogen preparation
  • Peptide synthesis: 2 different epitope peptides with 90% purity is synthesized by GenScript, 5 mg for each peptide. 2 mg of each free peptide will be delivered to the customer together with the final delivery.
  • Conjugation: The two peptides are conjugated to the carrier protein KLH for immunization.
14 days
Phase II: Immunization and cell fusion with MonoExpress™ protocol
  • Immunization: Balb/c mice or rats are immunized and boosted with MonoExpress™ immunization protocol.
  • Fusion: Electrofusion is performed and at least 10,000 clones can be obtained from each fusion. Additional fusions are available to reach our guarantee without extra charge.
45 days
Phase III: Screening and supernatant samples preparation for 1st delivery
  • Screening and isotyping: The hybridoma cells are expanded and screened by indirect ELISA. 4 clones (Including 1-3 sups for 2 epitopes) with best ELISA value and specificity to the free peptide are selected for isotyping.
  • 1st delivery: 2ml 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
  • Subcloning, expansion and cryopreservation: 4 clones are subcloned by limiting dilution, expanded and frozen.
  • Final Delivery: Two vials of cell culture for each cell line and 2 mg of each free peptide are delivered to the customer.
 30 days

MonoExpress™ Bronze (SC1660)   Fastest Custom Monoclonal Antibody Services
 
Description
Time Line
Phase I: Immunogen preparation
  • Peptide synthesis: 5 mg of peptide with 90% purity is synthesized. 2 mg of free peptide will be delivered to the customer together with the final delivery.
  • Conjugation: The peptide is conjugated to the carrier protein KLH for immunization.
14 days
Phase II: Immunization and cell fusion with MonoExpress™ protocol
  • Immunization: Balb/c mice or rats are immunized and boosted with MonoExpress™ immunization protocol.
  • Fusion: Electrofusion is performed and at least 10,000 clones can be obtained from each fusion. Additional fusions are available to reach our guarantee without extra charge.
45 days
Phase III: Screening and supernatant samples preparation for 1st delivery
  • Screening and isotyping: The hybridoma cells are expanded and screened by indirect ELISA. Up to 5 clones with best ELISA value and specificity to the free peptide are selected for isotyping.
  • 1st delivery: 2ml 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
  • Subcloning, expansion and cryopreservation: Up to 5 clones are subcloned by limiting dilution, expanded and frozen.
  • Final Delivery: Two vials of cell culture for each cell line and 2 mg of free peptide are delivered to the customer.
 30 days

MonoExpress™ Basic (SC1572)   Fastest Custom Monoclonal Antibody Services
 
Description
Time Line
Phase I: Immunization and cell fusion with MonoExpress™ protocol
  • Antigen: 1.5-2 mg of protein immunogen (purity >75%, >0.4 mg/ml) – provided by customer.
  • Immunization: Balb/c mice or rats are immunized and boosted with MonoExpress™ immunization protocol.
  • Fusion: Electrofusion is performed and at least 10,000 clones can be obtained from each fusion. Additional fusions are available to reach our guarantee without extra charge.
45 days
Phase II: Screening and supernatant samples preparation for 1st delivery
  • Screening and isotyping: Indirect ELISA is used as primary screening and positive clones are expanded for Western blot validation with protein immunogen. Up to 5 clones with best screening result are selected for isotyping.
  • 1st delivery: 2ml 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 III: Subcloning, expansion, cryopreservation, antibody production and final delivery
  • 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.
  • Final Delivery: Two vials of cell culture for each cell line and at least 2 mg purified antibody are delivered to the customer.
 30 days

MonoExpress™ Phospho-specific Antibody (SC1659)   Fastest Custom Monoclonal Antibody Services
 
Description
Time Line
Phase I: Immunogen preparation
  • Peptide synthesis: 5 mg of phospho-peptide (up to 15 residues) with >90% purity and 3 mg non-phospho-peptide with >70% purity are synthesized by GenScript. 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 for immunization.
28 days
Phase II: Immunization and cell fusion with MonoExpress™ protocol
  • Immunization: Balb/c mice or rats are immunized and boosted with MonoExpress™ immunization protocol.
  • Fusion: Electrofusion is performed and at least 10,000 clones can be obtained from each fusion. Additional fusions are available to reach our guarantee without extra charge.
45 days
Phase III: Screening and supernatant samples preparation for 1st delivery
  • Screening and isotyping: The hybridoma cells are screened by ELISA with phospho and non-phospho peptides and all positive clones are expanded into 24-well plates. Up to 5 clones with best ELISA value and specificity to phospho-peptide are selected for isotyping.
  • 1st delivery: 2ml 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
  • 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-phospho peptide < 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 free phospho-peptide and 2 mg of non-phospho peptide are delivered to the customer.
 30 days

Our customer service representatives are available 24 hours a day, Monday through Friday, to assist you.