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CUSTOM HYBRIDOMA DEVELOPMENT

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-Conjugate Service
-Hybridoma Development

OEM

 

PROCEDURE

 

Provide service to producing a hybridoma from your antigen includes the following steps:

 

STEP I

 

Immunization-5 certified balb/c mice would be immunized with customer¨s antigen. The antigen will be mixed with complete adjuvant for primary immunization and with incomplete adjuvant for booster immunization. An appropriate dose of the mixed antigen will be injected to different sites of mice. At around six weeks, following the primary immunization and first boost, the mice will be tested for antibody titer by ELISA assay. Only the mice that have ELISA titer at least 102-103 increased are used for the next step. Three to five days before the fusion, a qualified mouse will be selected for the pre-fusion boost.

 

The minimum immunization period is 8-12 weeks.

 

According to type of antigen and the antibody affinity requirement, we will choose different MaxMed Labs immunization procedures.

 

A minimum 2 mg of antigen is required for this step.

 

STEP II

 

Fusion & Screening-The spleenocytes from the immunized mouse will be prepared and fused with a myeloma cell line by appropriate ratio. The cells will be grown in HAT selection medium.

 

Screening and Selection-Supernatants from the resulting hybridoma clones are screened by ELISA against the antigen/carrier protein. The positive wells from the ELISA assays will be selected to be expanded for possible cloning.

 

The minimum period of this step is 3-5 weeks.

 

STEP III

 

Cloning-The positive wells will be selected for cloning by the limiting dilution method. Isolated colonies will be tested by ELISA again, and positives selected to be expanded for customer.

 

The minimum period of this step is 3-5 weeks.

 

A minimum 2 mg of antigen is required for step II and step III.

 

SCHEDULE AND PRICE

  • Whole period of one cycle is 16-20 weeks. More cycles maybe need for some projects.

  • Immunogen needed (customer provides):

                             Immunogen        4 mg

                             Hapten                 2 mg

                             Carrier Protein    2 mg

 

The immunogen purity should be more than 90%, without/with low concentration preservative and without any toxic chemical. The immunogen amount listed is the minimum required. The amount used may be very dependant on the number of positive clones that need screening.

  • Price: 20% of whole payment must be pre-deposit to open a project. It is not refund for any reason to stop the project.

 

Immunogen with carrier protein

(USD)

Immunogen without carrier protein

(USD)

Step I

1,500 (one antigen test)*

1,500

Step II

4,000 (one antigen test)*

3,000 (one antigen test)*

Step III

1,500

(one antigen test up to 3 clones)*^

1,500

(one antigen test up to 3 clones)*^

Total (USD)

$7,000

$6,000

*additional $500 in step I, additional $2,000 charge in step II and $1,000 in step III for every one more antigen screening

        ^additional $500 charge for every one more clone

 

Hybridoma cell line for customer

 

Final cell line(s) will be sent to customer in 15 ml sterile centrifuge tube(s) with 5-10 ml culture medium, and the cell density should be −2〜105/ml.

 

We only provide service and do not guaranty to get any specific clone that matches customers¨ particular criteria, such as sensitivity and specificity requirements. But normally, we can provide 1 positive cell line specific to the hapten and no cross to the carrier protein OR 2 positive cell lines to the immunogen. We do not ensure the cell lines have any specification to the additional screening antigens. We do not guarantee to get any cell line since the customers¨ immunogen problems.

 

Cell line QC Criteria

  • The supernatant for the test must be consistently cultured for 72 hours and the antibody titer must to be at least 10 times higher than the negative control (immunogen without carrier protein).
     

  • The cell culture with a density of at least 1-5x105 must be kept for 72 hours. Afterwards the supernatant is collected and tested. The specific antibody titer must to be at least 4 times higher than carrier protein (immunogen with carrier protein).
     

  • The cell lines have no general bacteria contamination. If customers need Mycloplasma tests. The test cost will be charged separately.

Cell line warranty

 

Please confirm the cell line to match our QC results between 90 days from the shipping date and sign on the Confirmation Form.  After 90 days, MaxMed Labs will not take any responsibility to reserve the cell lines.

 

Ascites and antibody production

 

Ascites Processing Service and Monoclonal Antibody Production (purification) Service are available. The quotation will be as required separately if necessary.

 

 

   
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