We provide researchers with high quality, reliable, custom antibody production services by recognizing that the humane care and use of the animals is essential to animal well-being as well as the quality of the services offered.
Since our inception in 1960, Major Academic Institutions, Nobel Prize Laureate sponsored laboratories, Major Pharmaceutical and Biotech companies have used Pocono Rabbit Farm and Laboratory (PRF&L) to help develop and produce their Custom Antibodies.
Be it Custom Antibodies to Cell Surface Receptors (e.g. GPCR), DNA, peptides or to highly conserved proteins, we can accommodate your single project or high throughput needs. Unlike some of our competitors, we do not subcontact out your antibody production. All work is done in our Canadensis, Pennsylvania facility. No need to go through separate vendors and establish various Purchase Orders to accomplish your goals, we are able to provide our clients with assistance from the selection and design of their immunogen through antibody purification. We have experience and technical staff to support the development of antibodies to a variety of antigen forms such as Native and Recombinant Proteins, Peptides, and Cells. Our goal is to provide you with High Quality “Functional” Custom Antibodies. We have been in the Antibody Production business for now over 50 years and are still family owned and managed. Our name and reputation stand behind every project.
28 DAY MIGHTY QUICK PROTOCOL PACKAGE
All 28 Day Mighty Quick Protocols include setup, prebleed, three immunizations (including immune stimulators), one large immune bleed, final bleed, and maintenance. This protocol can be extended if additional sera is required.
49 DAY MIGHTY QUICK PROTOCOL PACKAGE
All 49 Day Mighty Quick Protocols include setup, prebleed, four immunizations (including immune stimulators), four large immune bleeds, final bleed, and maintenance. This protocol can be extended if additional sera is required.
70 DAY POLYCLONAL ANTIBODY PRODUCTION
70 Day Antibody Production Package includes setup, prebleed, four immunizations, one small immune bleed, one large immune bleed, and maintenance. This protocol can be extended if additional sera is required. This protocol is based on the PRF&L Traditional 70 Protocol offered in the past.
84 DAY FUSION PROTEIN ANTIGEN POLYCLONAL ANTIBODY PRODUCTION
84 Day Fusion Protein Protocol, a favorite for many years at Pocono Rabbit Farm & Laboratory. This protocol uses an IV injection at day 56 as a way to improve the antibody. This package includes setup, prebleed, six immunizations, two large immune bleeds, and maintenance. This protocol can be extended if additional sera is required.
91 DAY POLYCLONAL ANTIBODY PRODUCTION
91 Day Antibody Production Package includes setup, prebleed, five immunizations, one small immune bleed, four large immune bleeds, and maintenance. All immunizations are subcutaneous for this package. This protocol can be extended if additional sera is required.
Our services includes cell line preparation (when provided with Mycoplasma free live cell line) for immunization into 5 Balb/c Mice, all necessary procedures and husbandry for a typical production run which is based on removing ascites fluid a maximum of 3 times per mouse followed by euthanasia and disposal. 3 to 10 ml per mouse can be anticipated.
Below are some considerations for using the ascites method for producing monoclonal antibodies.
CUSTOM HYBRIDOMA DEVELOPMENT AND MONOCLONAL ANTIBODY PRODUCTION
PHASE A. Immunization of BALB/c Mice and Serum Screening (approximately 8 weeks)
Eight mice are immunized with antigen(s) provided by customer. Antigen conjugation and purification services are available for an additional charge. At day 35, and later if necessary, mice are test bled and screened for titer by ELISA. If no acceptable response is found during this phase, $1500 will be retained for services up to this point and the remaining balance of deposit will be refunded.
PHASE B. Fusion and Screening of Hybridoma Supernatant (approximately an additional 8-10 weeks)
Upon successful completion of Phase A, selected mice will be euthanized and the spleens harvested. A fusion procedure will be performed and the cells will be distributed into six or eight multiwell plates (96 well) per spleen. Screening will be performed by ELISA using previously supplied antigen. Hybridoma lines from up to ten wells with the highest absorbency by ELISA will be amplified and the supernatants supplied to the investigator. If after the first fusion attempt no acceptable development is observed, a second fusion attempt will be made utilizing previously immunized mice remaining from Phase A. At this point if there is still no response, you the PI or PRF&L has the option to terminate at this point, the charge for completion of Phases A and B will be a total of $5000.
PHASE C. Cloning (approximately an additional 6-8 weeks totaling about 22-26 weeks)
One to five previously developed parental hybridomas will be selected, cloned and then screened by ELISA. Clones with desired titer and specificity will be expanded and cryopreserved. At least five clones will be provided and delivered to the customer depending on the stability of chosen hybridomas.
Total completion cost for phases A, B, & C is $7500 per antigen.
PHASE D. Custom Monoclonal Antibody Production by in vitro methods
Yield of approximately 50 to 100mg per batch
The customer's cell line is amplified using three liter tissue culture polyethylene bags. Monoclonal antibodies are isolated using a protein G column. Available only for IgG producing lines. - $1,450
Other protocols are available and the success with a certain protocol is dependent on the cell line.
No need to order your peptide from another vendor - PRF&L is able to meet your peptide synthesis needs.
Our basic package for antibody production is 10 to 15mg of peptide at 80 percent purity. Some investigators require higher purity or additional peptide, both of which are available.
For routine antibody production, 80 percent purity is sufficient. 85 percent, 90 percent, and 95 percent purity is available.
We are able to produce most common peptide modifications such as phosphorylation and acetylation.
Most peptides for antibody production ranges from 10 to 25 amino acids. If your peptide is over 25 amino acids, please contact us first for pricing as longer peptides can be more difficult to produce.
HPLC and Mass Spec data are available for peptides at no additional charge.
All peptides need to be conjugated to carrier protein. For most successful conjugation, the peptide should have a terminal cystine amino acid.
We are also able to conjugate peptides synthesized by other manufactures. Please make sure you tell us the peptide sequence.
We most commonly recommend conjugation to KLH, keyhole limpet hemocyanin. Other common conjugates BSA (bovine serum albumin) and OVA (ovalbumin).
Our standard conjugation package is to conjugate 4mg. 4mg of conjugated peptide is plenty to immunize several animals.
Unlike most of our competitors, PRF&L provides percent incorporation results on all conjugations we perform.
Free up your time and resources and allow PRF&L to verify specific antibody presence in your projects.
Antigen specific antibodies can be quantitated by Enzyme-Linked Immunosorbant Assay (ELISA). We use a 96 well format that gives an actual titer not just a + or - response and is preformed in duplicate.
Results are normalized back to the preimmune sera or previous bleed. Our staff can typically have the results back to you in electronic format before you would normally receive sera samples if we were to ship them for you to evaluate. This can also save you additional ship costs or simply act as an initial indicator of success prior to further more sensitive applications you may need to perform.
Protein Purification and Quantification Services
Biochemistry & Molecular Biology Services
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