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Monoclonal Antibody and Hybridoma Development Protocol

MonoExpress™ Premium (SC1711)

MonoExpress™ Basic (SC1717)

MonoExpress™ Gold (SC1710)

MonoExpress™ Silver (SC1661)

MonoExpress™ Bronze (SC1660)

MonoExpress™ Phospho-specific Antibody (SC1659)

Details and special requirements for hybridoma development

For a detailed description of a particular hybridoma development protocol, please choose your desired package above or browse our full suite of protocols below to find the one best suited to your needs. Should you have any questions, our technical specialists are available 24/7 to help.

If you have special requirements for your hybridoma development, please check out our fully Customized Monoclonal Antibody Services.

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.


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 hybridomas can be obtained from each fusion.
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 hybridomas are expanded. Ten hybridomas with best screening result are selected for isotyping.
  • 1st delivery: 10 ELISA (+) supernatants for evaluation in customer's own experimental system. Antibody Certificate of Analysis report is prepared with ELISA data and IgG concentration for each sample.
  • If a successful clone cannot be found after 1st delivery, customer has the option to receive an additional 10 supernatants*.
  • Cell line maintenance: The hybridomas 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: One hybridoma is 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 hybridoma selected by the customer.
  • Final Delivery: Up to 5 hybridoma cell lines of customer's choice+, 2 mg purified antibody from one chosen hybridoma, and 200µg immunogen protein are delivered to the customer.
30 days

+ Additional clones may be purchased at $399/clone.

* Read about our safety net policy

If a successful clone cannot be found, customer has the following options:

  1. Cancel the project after the first delivery of supernatants and only be charged $999 (applies to Premium (SC1711) and Basic (SC1717) packages).
  2. Purchase an additional 10 supernatants for $999. If a successful clone cannot be found after the second supernatant delivery, customer may choose to cancel the project and only be charged $999 plus the cost of the additional 10 supernatants, for a total of $1,999 (applies to Premium (SC1711) package only).

 
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 hybridomas can be obtained from each fusion. Additional fusions are available to reach our guarantee without extra charge.
45 days
Phase II: Screening and supernatant sample preparation for 1st delivery
  • Screening and isotyping: Indirect ELISA is used as primary screening and positive hybridomas are expanded for Western blot validation with protein immunogen. Five hybridomas with best screening result are selected for isotyping.
  • 1st delivery: 5 ELISA (+) supernatants for evaluation in customer's own experimental system*. 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: One hybridoma is 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 the hybridoma with best screening result.
  • Final Delivery: Up to 5 hybridoma cell lines of customer's choice+, 2 mg purified antibody from one chosen hybridoma.
30 days

+ Additional clones may be purchased at $399/clone.

* Read about our safety net policy

If a successful clone cannot be found, customer has the following options:

  1. Cancel the project after the first delivery of supernatants and only be charged $999 (applies to Premium (SC1711) and Basic (SC1717) packages).
  2. Purchase an additional 10 supernatants for $999. If a successful clone cannot be found after the second supernatant delivery, customer may choose to cancel the project and only be charged $999 plus the cost of the additional 10 supernatants, for a total of $1,999 (applies to Premium (SC1711) package only).

Description Time Line
Phase I: Immunogen preparation
  • Peptide synthesis: 3 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 hybridomas can be obtained from each fusion.
45 days
Phase III: Screening and isotyping 
  • Screening and isotyping: The hybridoma cells are expanded and screened by indirect ELISA. 6 hybridomas (2 derived from each peptide antigen) with best ELISA value and specificity to the free peptide are selected for isotyping.
Phase IV: Subcloning, expansion, cryopreservation, antibody production and final delivery
  • Subcloning, expansion and cryopreservation: Hybridomas are subcloned by limiting dilution, expanded and frozen.
  • Final Delivery: 6 hybridoma cell lines (2 clones/peptide); 6 supernatants++ at 5 ml/clone and 2 mg of each free peptide are delivered to the customer.
30 days

++ Purified mAb may be purchased at $399 for 2 mg purified mAb/clone.


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 hybridomas can be obtained from each fusion.
45 days
Phase III: Screening and isotyping 
  • Screening and isotyping: The hybridoma cells are expanded and screened by indirect ELISA. 4 hybridomas (2 derived from each peptide antigen) with best ELISA value and specificity to the free peptide are selected for isotyping.
Phase IV: Subcloning, expansion, cryopreservation, antibody production and final delivery
  • Subcloning, expansion and cryopreservation: Hybridomas are subcloned by limiting dilution, expanded and frozen.
  • Final Delivery: 4 hybridoma cell lines (2 clones/peptide); 4 supernatants++ at 5 ml/clone and 2 mg of each free peptide are delivered to the customer.
30 days

++ Purified mAb may be purchased at $399 for 2 mg purified mAb/clone.


Description Time Line
Phase I: Immunogen preparation
  • Peptide synthesis: 1 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 hybridomas can be obtained from each fusion.
45 days
Phase III: Screening and isotyping 
  • Screening and isotyping: The hybridoma cells are expanded and screened by indirect ELISA. 2 hybridomas (2 derived from each peptide antigen) with best ELISA value and specificity to the free peptide are selected for isotyping.
Phase IV: Subcloning, expansion, cryopreservation, antibody production and final delivery
  • Subcloning, expansion and cryopreservation: Hybridomas are subcloned by limiting dilution, expanded and frozen.
  • Final Delivery: 2 hybridoma cell lines (2 clones/peptide); 2 supernatants++ at 5 ml/clone and 2 mg of each free peptide are delivered to the customer.
30 days

++ Purified mAb may be purchased at $399 for 2 mg purified mAb/clone.


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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 hybridomas 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 hybridomas are expanded into 24-well plates. Up to 5 hybridomas with best ELISA value and specificity to phospho-peptide are selected for isotyping.
  • 1st delivery: 2ml supernatant samples for each hybridoma 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 hybridomas are subcloned by limiting dilution, expanded and frozen.
  • Antibody production: At least 2 mg purified antibody is produced for the hybridoma with best screening result. The cross reactivity with non-phospho peptide < 10% for the purified antibody.
  • Final Delivery: Two vials of cell culture for hybridoma, 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

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