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Rockland has extensive experience producing monoclonal antibodies and we offer services that are highly customized to your specific requirements. We provide a comprehensive service which includes all steps of monoclonal antibody production from peptide synthesis to ascites production.
Below is a general procedure for production of Monoclonal Antibodies. As each project is unique and experimental needs differ, we at Rockland have the knowledge and experience to help you design the most scientifically sound approach to achieve your goals.
Please inquire for additional information, methodology or a written price quotation contact our Custom Services Department.
| HYBRIDOMA DEVELOPMENT |
|
Service |
Code |
Price |
| Phase 1 (Steps 1-3) |
MAB-001 |
inquire |
 Step 1: Immunization of 5 Balb/c mice* - In general we begin with a 50 microgram priming dose followed by several boosting doses of 50 microgram each. This regime will continue until a sufficient antiserum titer is achieved.
Step 2: Screening-Test bleeds are routinely performed and evaluated by Rockland's staff using a direct binding ELISA. Optimal ELISA conditions are developed to ensure the proper evaluation of future clones. In addition test bleeds can be sent to your lab for evaluation in other assays.
Step 3: Evaluation-Data is evaluated jointly with Rockland staff to determine the performance of antiserum at this point. Antiserum titers and specific assay performance must be acceptable to continue the project. |
| Phase 2 (Steps 4-5) |
MAB-002 |
inquire |
| Step 4: Fusion-A splenocyte fusion is performed on the mice (2) which responsed the best to the immunizations as determined by ELISA or another assay. The lymphocytes are then fused to an Sp2/0-Ag14 cell line using an optimized PEG mediated fusion protocol.
Step 5: Expansion & Screening-The fusion is plated into a stack of ten to twenty 96 well plates. Plates are monitored for growth and fed weekly. Wells with cell growth are screened by ELISA in 2 to 3 weeks. |
| Phase 3 (Steps 6-7) |
MAB-003 |
inquire |
| Step 6: Subcloning-Cells from five parental positive wells are harvested and subcloned by limiting dilution.
Step 7: Screening & Expansion-Subclones are screened by ELISA, positives expanded, and screened again. Five sublcones will be expanded, frozen and stored for up to six months. |
Total Cost for Phase I through III (Steps 1 through 7) .......... inquire
Time: Approximately 4-6 months depending on the response to the Antigen
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HYBRIDOMA DEVELOPMENT
& Additional Services |
|
Service |
Code |
Price |
Additional Colony Expansions
(in addition to the five already included) |
MAB-004 |
inquire |
| Step 6 of Rockland's complete service includes up to 5 positive parental clones. Additional subcloning and expansion can be performed. |
| Liquid Nitrogen Storage |
MAB-005 |
inquire |
| Precious clones can be maintained in liquid nitrogen in our monitored facility. |
| Mycoplasma Test for Cell Lines |
MAB-012 |
inquire |
| The hybridoma cell lines developed by Rockland or from other sources can be tested for any Mycoplasma contamination. |
| Bioprocessing |
MAB-090 |
inquire |
| Antibodies produced from Hybridoma cell lines can be characterized for epitope mapping, native protein reaction, cross reaction and titration using immunoprecipitation, immunoblotting and various ELISA methods. Please Inquire for additional details. |
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| ASCITES PRODUCTION |
|
Service |
Code |
Price |
| Ascites Production in Balb/c Mice (5) |
CUST25 |
inquire |
| Ascites Production in Balb/c Mice (10) |
CUST26 |
inquire |
| Ascites Production in Balb/c Mice (20) |
CUST27 |
inquire |
| Ascites Production in Balb/c Mice (50) |
CUST71 |
inquire |
| Ascites Production in Balb/c Mice (100) |
CUST72 |
inquire |
| Ascites Production in Balb/c Mice (200) |
CUST73 |
inquire |
| Ascites is produced in Balb/c strain mice. Optimized methods are used to generate tumors and accumulate ascitic fluid, including the quantity of cells and volume of inocula. These methods generate high yields of antibody that can be be further processed or characterized depending on your particular requirements. |
| Ascites Prod. using Immunodeficient Mice (5) |
CUST51 |
inquire |
| Ascites Prod. using Immunodeficient Mice (10) |
CUST52 |
inquire |
| Ascites Prod. using Immunodeficient Mice (20) |
CUST53 |
inquire |
| Ascites Prod. using Immunodeficient Mice (50) |
CUST74 |
inquire |
| Ascites Prod. using Immunodeficient Mice (100) |
CUST75 |
inquire |
| Ascites Prod. using Immunodeficient Mice (200) |
CUST76 |
inquire |
| Immunodeficient mouse strains (SCID, nude or RAG -/-) are used for ascites production when fusion partners are derived from an animal other than the Balb/c mouse. Please consult with our technical staff for additional information regarding this service. |
Please inquire for larger quantities of mice for ascites production.
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Please inquire for larger volumes or technologies not listed above.
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| CELL CULTURE SERVICES |
|
Service |
Code |
Price |
| Adherent Cell Cultures |
MAB-080 |
inquire |
| Small and large scale cultures can be produced. Typically flasks or cell factories are used for production |
| Suspension Cell Cultures |
MAB-082 |
inquire |
| Small and large scale cultures can be produced. typically any method listed for in-vitro antibody production can be used. Please inquire if you require a method not listed. |
| Production of Bioactive Molecules |
MAB-083 |
inquire |
| In Vitro methods are widely utilized for the production of some bioactive molecules. Rockland uses tissue culture methods and other technologies for the production of recombinant proteins, hormones, growth factors and viral products in both small and large scale quantities. Please inquire for specific details on this service. |
| Mammalian Primary Cell Line Derivation |
MAB-009 |
inquire |
| Primary cell line derivation can be performed on a variety of tissue types from several different species. Cell lines include, but are not limited to, embryonic stem cell culture, lymphocytes and splenocytes. Please inquire for specific details on this service. |
| Cell Lysate Service |
MAB-081 |
inquire |
| Cell pellets generated by Rockland or supplied to Rockland can be processed to produce total protein lysates prepared for SDS-PAGE electrophoresis. Chromosomal DNA is sheared and clarified supernatant containing total protein is supplied in SDS-PAGE buffer with DTT and bromphenol blue. Please inquire for nuclear and subcellular fractionations. |
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Tissue Culture Production of Monoclonal Antibodies
edited last: March 23, 2007
WebMaster |
