ProSci Incorporated is a privately-held biotech firm located in San Diego, California. A leader in its field, ProSci focuses its energies in Antibody Research Products, Custom Antibody / Immunochemistry Services and Research Programs. By combining its specializations in each of these disciplines, ProSci is able to provide its customers with practical experience, innovative products, competitive pricing, high-quality research tools, and customized services. In line with its research product offerings, ProSci offers custom polyclonal and monoclonal antibody production services using in house laboratories. To enhance the value and accuracy of these custom produced antibodies, ProSci provides a complete spectrum of immunochemistry services. These include antigen design assistance, antibody characterization, peptide synthesis, protein conjugation, IgG and affinity purification, ELISA, Western Blot Mass Spectrometry, and Immunohistochemistry. In conjunction with its strengths in the antibody and reagent market, ProSci Incorporated actively conducts research in a variety of fields. Most notably, ProSci is testing a novel tumor marker for diagnostic and therapeutics applications. In addition, ProSci is performing research in fields such as AIDS vaccine development, stem cells, proteomics and bioterrorism. This capacity to innovate again becomes a service for the customer, as it allows ProSci to offer contract research alliances with outside institutions. In such situations, ProSci provides large-scale customized immunochemistry services tailored to the needs of the institution, thereby permitting the reduction of research costs, the outsourcing of immunochemistry services and the incorporation of high-grade antibody products into research projects. In the end, the use of these services allows the researcher to reduce research expenses, outsource specialized immunochemistry services and incorporate quality products into the research process. Moreover, our own experience in using these products and services for research purposes enables us to provide the customer with detailed and relevant advice concerning the use of antibody and immunochemistry technologies. To this end, we invite you to explore our custom antibody and immunochemistry service offerings.
ProSci is proud to offer Single Domain Antibody Production also known as Camelid, VHH, and Nanobody Production. Each project includes 5 phases as listed below.
Single Domain Antibody(sdAb) Development process
Phase I: Immunization
Immunization of 1 llama on standard 12-week protocol with 4 immunizations and 3 bleeds
ELISA using each bleed screened against antigen
Phase II: PBMC Isolation
PBMC's isolated and preserved from 100mL blood collected at weeks 8 and 12. Half of PBMCs are immediately lysed and cryopreserved for library construction.
Phase III: Library Construction
RNA isolation from Phase II PBMCs(customer's choice of bleed, 2 or 3, >5 x 10^7 cells or a pool using >5 x 10^7 cells from each bleed)
sdAb library construction
Phase IV: Library Screening
Panning of phage library against antigen
Screening by indirect ELISA
Selection and sequencing of high affinity binders
Relative affinity determination by ELISA
100 µg of up to 6 final clones (clones from distinct sequence families)
Additional screening data (if requested; additional charge)
Phase V : Production (Optional)
Expression and purification of additional quantities of antibody
Polyclonal antibodies are a mixture of immunoglobulins that recognize different epitopes on a specific antigen. ProSci produces polyclonal antibodies in rabbits, mice, rats, chickens, goats, or llamas. The antibodies are isolated from the serum of mammalian hosts or from the yolks of chicken eggs. Serum can be used directly or further purified, but yolks are further purified prior to use to remove the lipids that can interfere in assays. We offer a standard antibody production protocols that balance fast turnaround time with quality for each host species. We offer packages for client-provided antigens, single-peptide projects, peptide cocktail projects, and phospho-peptide projects. An overview of our standard packages includes: - **Serum Package** - *Antibody Production using Client-Provided Antigen* - Immunization protocol in desired host species - **Peptide Package** - *Antibody Production using peptide synthesized by ProSci* - Synthesis of peptide up to 15 residues (5 mg for customer) - Conjugation of peptide to carrier protein - Immunization of conjugate into desired host species - ELISA on Bleed #1 to verify titers using unconjugated peptide - **Purified Package** - Antibody Production using peptide synthesized by ProSci - Synthesis of peptide up to 15 residues (5 mg for customer) - Conjugation of peptides to carrier protein - Immunization of conjugate into desired host species - ELISA on Bleed #1 to verify titers using unconjugated peptide - Immuno-Affinity Purification of up to 25 mL serum on last scheduled bleed (includes re-usable column) Upon completion of the initial immunization protocol included in the above packages there are several options to select from: - Termination - Smaller species are euthanized and goats and llamas are retired without any additional services - Monthly Extension - 1 immunization - 2 bleeds - Exsanguination - Add-on service for chickens only; included in mouse, rabbits, rat, and guinea pig schedule - Chicken: single bleed that yields ~15 mL serum Note: Mice production bleeds are terminal bleeds; if extension is desired it must be requested prior to the terminal bleed. We offer extensive antibody services to add to each package and welcome custom schedules to create a custom package for your antibody needs. *Peptide Cocktail Polyclonal Antibody Packages* include all of the services in the Peptide Package and Purified Package respectively, but also include the synthesis, conjugation, and immunization of your choice of 2 peptides into one set of animals with the option to upgrade to 3 peptides. These packages increase the likelihood of a peptide-based antibody working for multiple applications as it doubles or triples the available number of epitopes for the animals’ immune systems to respond against. It also helps to reduce overall cost by minimizing the number of animals needed for a particular project. *Phospho-Specific Polyclonal Antibody Packages* include all of the services in the Peptide Package and Purified Package respectively, but the peptide immunogen includes a phosphorylated amino acid. The Purified Package also offers synthesis of the corresponding non-phospho peptide sequence, used for isolating both the phospho and non-phospho antibodies for your target protein. Other modification-specific custom antibody services available upon request. *Guaranteed Polyclonal Antibody Packages* include all of the services in the Serum Package and Purified Package but with additional guarantees. The Guaranteed Serum Package includes a titer guarantee of >1:50,000 for a protein antigen provided by you and the Guaranteed Purified Package guarantee an immuno-affinity purified antibody for use in Western Blotting. The Guaranteed Packages require prior approval before acceptance and initiation.
As part of our additional services, ProSci purifies antibodies from serum, ascites fluid, culture medium, or yolks using a variety of purification procedures. These procedures include protein A or G, immuno-affinity, or IgY purification.
Protein A or G:
These proteins isolate total mammalian IgG from serum, ascites fluid, or cell culture medium. Selecting Protein A or Protein G depends on the sample’s species and antibody isotype. Protein A or G is not recommended for peptide antigens as antibodies against the large carrier protein will also be isolated. This method is best used with antiserum produced using protein antigens larger than 10 kDa and for purifying monoclonal antibodies from ascites or supernatant.
This method isolates antigen-specific antibodies using a column constructed by coupling an antigen to a gel matrix. It is best used for isolating peptide-specific polyclonal antibodies from serum, but can also be used with protein antigens and for purifying monoclonal antibodies.
This method isolates total IgY from yolks. Chicken IgY is the functional equivalent of mammalian IgG but is primarily isolated from egg yolks rather than serum. This process is performed by removing lipids first and then precipitating antibodies from the resulting solution. The total IgY can then be used “as-is” or further purified using an immuno-affinity column.
Antigen design assistance for peptide antigens. If you can provide us with the sequence (or protein accession number), our scientists will analyze your protein and select the regions which offer the best chance of eliciting an immune response for a variety of applications, including western blot, immunohistochemistry, immunocytochemistry, and immunoprecipitation. We base our sequence analysis on several factors:
If you choose to use a sequence we have designed or approved for one of our standard antibody production packages utilizing peptide synthesis, we will guarantee the immune response against the peptide antigen. If no titer is observed, ProSci will re-immunize a new set of animals. If a titer against the antigen is still not present, ProSci will re-evaluate the protein of interest and synthesize a new peptide at no cost. ProSci will continue to work on your project until a successful immune response is observed. Guarantee does not apply to modification specific or peptide cocktail packages or host species other than rabbit.
As part of our additional services, ProSci offers custom peptide synthesis. We strive to provide high quality production with quick turnaround time for our customers.
Peptide Synthesis Capabilities:
70% is typically considered immunograde and is enough for making polyclonal antibodies. However, HPLC to >90% is recommended for peptides with modified amino acids and monoclonal antibody development. ProSci can provide peptides from crude up to >98% purity.
We also offer complimentary peptide-antigen design for our monoclonal and polyclonal packages.
Monoclonal antibodies are secreted by an immortal cell line typically derived from spleen cells fused with a myeloma cell line. They recognize only one epitope on an antigen. Once a single, stable clone is isolated, the antibodies can be made either in vitro or in vivo with ascites or cell culture supernatant.
ProSci helps develop custom monoclonal antibodies and understands researchers have varying needs for levels of involvement, depending on the target. We offer custom monoclonal antibody development packages for both protein and peptide antigens and add-on services for customization.
Our screening options include:
ProSci’s fusion success rate for custom monoclonal antibody development is greater than 95% using techniques developed in-house. All work is performed by skilled technicians on premises in our San Diego, California laboratories. We can screen against multiple antigens (additional charges apply) to develop the specific clone you need. ProSci has repeatedly proven our abilities to produce hybridomas against difficult targets.
We use 5 BALB/c mice as a production strain for hybridoma development. BALB/c is a very common species used throughout the industry for monoclonal antibody development due to their large spleen to body size ratio. This yields a higher number of splenocytes at fusion.
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