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Custom Immunology Services
Technical Tips for Successful Antibody Production |
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Project Planning
Immunogen Preparation
Shipping
Study Conclusion
Project Planning
- Peptides
- Assistance for peptide design based on antigenicity is FREE. Have sequence available or forward for review.
- Standard peptide purity is 80%. If you require higher purity, please specify this request on your order.
- A cysteine is typically added for conjugation. Please provide location of cysteines in your sequence if we are conjugating your peptide.
- Phosphorylated and MAP peptides are available.
- Peptide is delivered as a lyophilized powder.
- All peptides are delivered with HPLC and mass spec analysis.
- Peptide synthesis typically takes 2-4 weeks.
- Higher purity of immunogen will provide a more specific antibody response.
- Pre-screening, performed by your lab or ours, gives you the option to screen a chosen number of animals for any background. You may then select or release these animals. Please provide screening antigen for Covance to complete.
- Specific background concerns should be discussed with Covance. Diet and other factors can be altered to decrease certain background responses.
- Our standard schedule of events (multiple species) are 118 days and have proven to provide the highest yield and highest quality antibodies over other available shorter schedule of events. You may provide a custom schedule of event or use our accelerated schedule of event option.
- Due to the variant nature of immune response from animal to animal, it is recommended that a minimum of 2-5 animals be used per immunogen.
- Chickens are an excellent host for highly homologous mammalian proteins/sequences and will decrease incidence of cross reactivity.
- Other species available include various SPF mice, rats, guinea pigs, rabbits, goats, sheep, swine, bovine, and equine.
- It is recommended that your lab or ours monitor titer of antigen specific antibody presence by ELISA. ELISA screening can be performed on any pre, test, production, or terminal bleed. Proactive steps can be taken throughout the study to increase response to antigen.
- IgG purification followed by antigen specific affinity purification will provide the most pure serum. A safety net procedure should be in place to assure acceptable recovery levels of antibody.
- Call your scientific support team at Covance as needed for assistance on any aspect of your study.
Immunogen Preparation
- Immunogen used for antibody production may be in a buffer, lyophilized, or in gel. We cannot accept immunogens containing certain components. For specific questions regarding your buffer, please feel free to contact one of our Client Care Specialists at 717-336-4921 or 800-345-4114.
- The recommended concentration of immunogen is 1.0 mg/mL or greater and the recommended buffer is PBS. Please label vials with concentration and volume.
- If your immunogen is insoluble, you may send it in gel. The gel may be stained, but should be rinsed with water to remove any residual chemicals. Gel should be placed in a tube with no buffer, should remain pliable, and does not have to be crushed. We will prepare the emulsion for injection. Gel cannot be used for ELISA; a soluble antigen is required.
- Fusion proteins can be used for the immunization phase, but the cleaved protein or protein with an alternate carrier will be needed for ELISA screening. Conjugated peptides can and should be used for immunization as well, but unconjugated peptide or peptide conjugated to an alternate carrier will be needed for screening. An additional 100 ug of antigen per animal suitable for plate coating is required for ELISA screening.
- It is recommended that you aliquot your immunogen for each injection to minimize freeze-thaw cycles. There should be enough material in each vial to dose all animals to be immunized (on a date) with that specific immunogen. Any excess will be maintained and returned to you at your request at the termination of the study.
Shipping
- ALL orders must have a completed order form including as much of the requested information as is applicable. Send additional information as needed to clarify. Refer to assigned quote number when sending any material to Covance. If the study is ongoing, please include a note with your name, institution, and animal numbers for identification.
- Covance order forms contain two pages. Page one is for project / client information and the second page is an Assurance statement which must be signed. Please complete and include both pages to initiate your project.
- When shipping immunogen on dry ice, do not seal any dry ice into a container such as a screw-capped tube. This can explode upon removal.
- Do not use thin glass vials for storage of your immunogen. They have a tendency to break during freezing and especially during shipping. Please label all tubes with immunogen name, volume, and concentration.
- Shipping and packaging by Fed Ex (or best way) averages $47 per shipment in the United States. Contact us for international rates.
- Please be aware of your country's policies on shipment of biological materials and dry ice.
- You may modify your shipping schedule and/or combine shipments.
Study Conclusion
- A study will be concluded only with consent of the investigator. Call your scientific support team at Covance to discuss study conclusion options.
- If permission is not given to conclude the study, maintenance fees will accrue.
- Storage of Antibodies:
- Avoid repeated freezing and thawing of antibodies.
- Aliquot antibodies in volumes that will be used within a reasonable time, usually one to two months.
- Keep aliquots frozen except for volume at 2-8°C. Carefully label the aliquoted antibodies and document thaw date.
- Store antibodies undiluted. The more dilute the antibody, the less stable. Do not dilute antibodies until you are ready to use them.
Download needed order forms. We'll process your order immediately.
Our Shipping Address
Attn: Immunology Services
465 Swampbridge Road
Denver, PA 17517
tel (800) 345-4114
tel (717) 336-4921
fax (717) 336-3481
AAALAC International Accredited, GLP Studies, OLAW Assured, USDA Research Registered
Document Revision Date: 8/28/07 |
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