POLYCLONAL ANTIBODY DEVELOPMENT

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1 POLYCLONAL ANTIBODY DEVELOPMENT Abcore is committed to excellence, and to responding to the changing needs of our customers. We have developed polyclonal antibodies for biotechnology and pharmaceutical companies, research institutions, and universities throughout the United States and internationally. The protocols and methods used to produce polyclonal antibodies are specifically suited to small scale and large scale requirements. The highest quality is achieved through careful and diligent attention to the health of our animals. Healthy animals make better antibodies. When you develop a custom antibody protocol with Abcore we guarantee to provide quality immunological services. We use New Zealand White females rabbits, bred on our farm, and adult laying leghorn chickens, Boer goats, and sheep. All of our animals are cared for by qualified personnel to ensure that you are receiving the highest quality service. All antibody work is done by Abcore personal we do not subcontract. Our facilities are USDA registered #93-R-0541 and maintained by our IACUC committee to insure all regulations are being followed. We keep our customers in touch of their project progress every step of the way by updates of project progress and are available to answer questions at any time. Attached you will find 1) Standard Protocols for Rabbits, Chickens, Goats, and Sheep 2) Antigen requirements 3) Project outline If you need additional information, a custom price quote for multiple animals, or are ready to order services, please contact an Abcore. We look forward to working with you. Sincerely, Abcore Staff 405 Maple St., Suite A106, Ramona, CA T F P a g e 1

2 RABBIT POLYCLONAL PRICING STANDARD PROTOCOL (Two Rabbits) $ STANDARD RABBIT PROTOCOL (60 Days) Week Week 0 Week 2 Week 4 Week 5 Week 6 Week 7 Week 8 ACTION Pre-immunization bleed only upon request Immunize with.25mg of antigen mixed with Complete Freund's Adjuvant Immunize with.2mg of antigen mixed with Incomplete Freund's Adjuvant Immunize with.2mg of antigen mixed with Incomplete Freund's Adjuvant Production bleed - ~20-25ml of serum Immunize with.2mg of antigen mixed with Incomplete Freund's Adjuvant Production bleed - ~20-25ml of serum Production bleed - ~20-25ml of serum ELISA or other testing requested by client Shipment or Purification of Serum Antigen requirement: 1.7 mgs per 2-rabbit protocol for Immunizations Pre-immunization bleeds average 5 ml serum only if requested 1 Bleed ELISA is done at any time during the project at no charge if you want more than one ELISA pricing is below. Additional antigen is required if you request ELISA testing. Average yield of serum from the Standard Protocol is ml. Once you receive your bleeds you have 2 weeks per diem no charge after the 2 weeks per diem will start if you want to continue project. ADDITIONAL ITEMS if Project is extended past 60 Days PER DIEM BOARD AND CARE PRODUCTION BLEEDS (Avg ml Serum) SQ/IM/IP/ID/IV IMMUNIZATIONS TEST EXSANGUINATION/Bleed BLEEDS (Avg. 7-10ml Out Serum ADDITIONAL ELISA ASSAY AFFINITY PURIFICATION (additional antigen needed) Protein G Purification $1.50/Day/Animal $17.00/Procedure/Animal $13.00/Procedure/Animal $14.85/Procedure/Animal $45.00/Animal $75.00/Plate $500.00/25.0ml >25.0ml 10.00/ml $ ml >50ml $10.00/ml 405 Maple St., Suite A106, Ramona, CA T F P a g e 2

3 PLEASE INQUIRE FOR DISCOUNTS ON MULTIPLE ANIMALS GOAT/SHEEP POLYCLONAL PRICING STANDARD PROTOCOL (One Goat) $ Week STANDARD GOAT/SHEEP PROTOCOL (98 Days) ACTION Pre-immunization bleed only upon request Week 0 Immunize with.5mg of antigen mixed with Complete Freund's Week 2 Adjuvant Immunize with.5mg of antigen mixed with Incomplete Freund's Week 4 Adjuvant Immunize with.5mg of antigen mixed with Incomplete Freund's Week 6 Production bleed - ~ ml of serum Week 8 Immunize with.5mg of antigen mixed with Incomplete Freund's Week 10 Adjuvant Production bleed - ~ ml of serum Week 12 Immunize with.5mg of antigen mixed with Incomplete Freund's Week 14 Adjuvant Production bleed - ~ ml of serum Adjuvant Antigen requirement: 2.5 mgs per 1 goat. Pre-immunization bleeds average 5 ml serum only if requested 1 Bleed ELISA is done at any time during the project at no charge if you want more than one ELISA pricing is below. Additional antigen is required if you request ELISA testing. Average yield of serum from the Standard Protocol is 750ml serum Once you receive your bleeds you have 2 weeks per diem no charge after the 2 weeks per diem will start if you want to continue project. ADDITIONAL ITEMS if Project is extended past 98 Days PER DIEM BOARD AND CARE PRODUCTION BLEEDS (Avg ml Serum) SQ/IM/IP/ID/IV IMMUNIZATIONS TEST ADDITIONAL BLEEDS ELISA (Avg. 7-10ml ASSAY Serum AFFINITY PURIFICATION(additional antigen needed) Protein G Purification $5.00/Day/Animal $50.00/Procedure/Animal $15.00/Procedure/Animal $14.85/Procedure/Animal $75.00/Plate $500.00/25.0ml >25.0ml 10.00/ml $ ml >50ml $10.00/ml 405 Maple St., Suite A106, Ramona, CA T F P a g e 3

4 LLama POLYCLONAL PRICING STANDARD PROTOCOL (One Llama) $ Week STANDARD Llama PROTOCOL (91 Days) ACTION Pre-immunization bleed only upon request Week 0 Immunize with.5mg of antigen mixed with Complete Freund's Week 2 Adjuvant Immunize with.5mg of antigen mixed with Incomplete Freund's Week 4 Adjuvant Immunize with.5mg of antigen mixed with Incomplete Freund's Week 6 Production bleed - ~ ml of serum/500ml whole blood Week 8 Immunize with.5mg of antigen mixed with Incomplete Freund's Week 10 Adjuvant Production bleed - ~ ml of serum/500ml whole blood Week 12 Immunize with.5mg of antigen mixed with Incomplete Freund's Week 14 Adjuvant Production bleed - ~ ml of serum/500ml whole blood Adjuvant Antigen requirement: 2.5 mgs per 1 Llama. Pre-immunization bleeds average 5 ml serum only if requested 1 Bleed ELISA is done at any time during the project at no charge if you want more than one ELISA pricing is below. Additional antigen is required if you request ELISA testing. Average yield of serum from the Standard Protocol is 750ml serum Once you receive your bleeds you have 2 weeks per diem no charge after the 2 weeks per diem will start if you want to continue project. ADDITIONAL ITEMS if Project is extended past 98 Days PER DIEM BOARD AND CARE PRODUCTION BLEEDS (Avg ml Serum) SQ/IM/IP/ID/IV IMMUNIZATIONS TEST ADDITIONAL BLEEDS ELISA (Avg. 7-10ml ASSAY Serum AFFINITY PURIFICATION(additional antigen needed) Protein G Purification $10.50/Day/Animal $450.00/Procedure/Animal $32.50/Procedure/Animal $14.85/Procedure/Animal $75.00/Plate $500.00/25.0ml >25.0ml 10.00/ml $ ml >50ml $10.00/ml PLEASE INQUIRE FOR DISCOUNTS ON MULTIPLE ANIMALS 405 Maple St., Suite A106, Ramona, CA T F P a g e 4

5 CHICKEN POLYCLONAL PRICING STANDARD PROTOCOL (2 Chickens) $ STANDARD CHICKEN PROTOCOL (56 Days) Week Week 0 Week 2 Week 4 Week 5 Week 7 Week 8 ACTION Pre-immunization egg collection Immunize with.25mg of antigen mixed with Complete Freund's Adjuvant Immunize with.15mg of antigen mixed with Incomplete Freund's Adjuvant Immunize with.15mg of antigen mixed with Incomplete Freund's Adjuvant Start Egg Collection Immunize with.15mg of antigen mixed with Incomplete Freund's Adjuvant End Egg Collection Antigen for a 2 chicken project is 1.4mg Yield is ~12-15 Eggs per chicken IgY yield is approximately 150mgs/yolk Once you receive your eggs/igy you have 2 weeks per diem no charge after the 2 weeks per diem will start if you want to continue project. Yolk isolation is available for $1.00/egg and will be shipped in PBS containing 0.1% NaN3 we can also intact eggs. ADDITIONAL ITEMS if Project is extended past 56 Days PER DIEM BOARD AND CARE /Includes egg collection IMMUNIZATIONS IgY PURIFICATION ELISA ASSAY on purified material AFFINITY PURIFICATION on IgY purified material (additional antigen needed) $2.00/Day/Animal $10.00/Procedure/Animal $150.00/1 Egg $75.00/Plate $500.00/25.0ml >25.0ml 10.00/ml 405 Maple St., Suite A106, Ramona, CA T F P a g e 5

6 Contact / shipping Information Company/Institution Contact Address City State Zip Phone Fax Purchase Order Number: CC# Expiration Date Name on Card: Billing Information Same as Contact/Shipping Company/Institution Attention Address City State Zip Phone Fax Purchase Order Number: CC# Expiration Date Name on Card: 405 Maple St., Suite A106, Ramona, CA T F P a g e 6

7 IMMUNIZATION REQUIREMENTS Number of Animals: Species of Animal: Rabbits Chickens Goat Sheep Rabbit Project Type: Pre Bleed Yes No Standard Rabbit Custom Rabbit (If custom attach protocol) Chicken : Chicken Project Type: Yolk Isolation 1 yolk/tube (Additional Charge) IgY Purification(Additional Charge) Goat/Sheep Standard : Chicken Custom Chicken Program 1 Program 2 Standard Goat /Sheep Custom Goat /Sheep(If custom attach protocol) ****Please attach any custom protocols**** ANTIGEN INFORMATION Antigen Name: Peptide Sequence: Peptide Protein Other Explain: % of Purity: Antigen Size : Molecular Weight Antigen Form: Liquid Dry/Lyophilized Frozen Gel Slices Number of Vials: Buffer antigen is in: Concentration: Volume: ml Dry mg Disposition of Antigen: Return with final shipment Discard Antigen Storage: ** Abcore cannot accept antigens that are pathogenic, toxic, and/or carcinogenic ** 405 Maple St., Suite A106, Ramona, CA T F P a g e 7

8 Approval is needed from Abcore prior to shipment of antigen. We cannot accept any toxic, radioactive, human derived, live bacteria, virus, and fungus. Does antigen contain radioactive material? Is antigen known to be toxic? Is antigen from, or related to human derived material? Does antigen contain live bacteria, virus, fungus, or protozoan? Does antigen contain killed or inactivated bacteria, virus, fungus, or protozoan? No No No No No ANTIGEN REQUIREMENTS PER ANIMAL: To avoid delays, please send no less than the minimum amount required Rabbit(2) 1.7mg Chicken(2) 1.4 mg Goat/Sheep(1) 2.5 mg Additional Antigen or free peptide will be required for: ELISA titration 1.0 mg Sm Affinity Purification <30mL 10mg additional antigen required Lg Affinity Purification mg additional antigen required 200 X-Lg Affinity Purification mg per each additional 100mL 405 Maple St., Suite A106, Ramona, CA T F P a g e 8

9 CONJUGATION OF ANTIGEN Is Antigen conjugated to a carrier Yes No What Carrier: **Abcore recommends peptides are conjugated to KLH for immunization and a carrier other than KLH for testing most common is BSA for testing** If antigen is not conjugated is it required? Yes No KLH BSA THY Other: Free Reactive Group: Cysteine Carboxyl Amino Sequence ANALYSIS REQUIREMENTS ELISA Titration on Samples: Yes No (1 included for no additional charge) If Yes At End of Project Other Western Blot: Yes No (additional charge) Blot against: PURIFICATION/ADDITIONAL PROCESSING (optional) IF NO PURIFICATION IS REQUIRED PROCEED TO PACKAGING SECTION Would you like the antibody purified? Yes No All Volume: ml Additional Testing : After each bleed Assay Samples: Method of Purification: Protein G Affinity *** Additional 10mg of antigen required for Affinity Purification) Would you like 0.1% NaN3 added to purified antibody? Yes No Do you have a preference for final delivery buffer (default PBS ph 7.4)? Yes No Custom Do you have a preference for concentration of final antibody? Yes No mg/ml Would you like 50% glycerol added: Yes No *** All Antibody is 0.2um Sterile filtered prior to packaging *** Yes No 405 Maple St., Suite A106, Ramona, CA T F P a g e 9

10 CONJUGATION OF PURIFIED ANTIBODY Do you want your polyclonal antibody conjugated: Yes No HRP AP Biotin FITC Amount: PACKAGING Final Packaging: Bulk aliquot Aliquot size: MG/Vial Multiple aliquots, Aliquot size: ML/Vial Do you want 0.1% NaN3 added to your Sera : Yes Shipping conditions: No Remaining Ice Packs Antigen: Dry Ice Return With Final Shipment Discard Retain (6 Months) ***Dry Ice Charges Apply*** SHIPPING METHOD Shipping Schedule: End of Project After Each Bleed Custom UPS FEDEX Prepay and add to invoice UPS/ FEDEX Account Number: Priority (Additional Charges Apply) Standard Please note: Animals will not be terminated without specific written instructions from the client. You will be contacted after your 2 week waiting period to determine if you want your animals terminated or extended. 405 Maple St., Suite A106, Ramona, CA T F P a g e 10

11 TERMS AND CONDITIONS PRICES Prices of services do not include freight and packing, VAT or import duties where these are applicable. Prices and other information provided are subject to change without notice. DELIVERY Orders received by Abcore will be confirmed in writing including a project schedule with immunization, bleeding and shipping dates. To ensure quality and stability of the product and minimize freight costs, Abcore and its customers should agree on packaging and best way of shipping. ABCORE LIMITED WARRANTY All preparations are for research or further manufacturing use only. They are not intended for any human or animal therapeutic use. Abcore provides services making custom-designed antibodies; therefore Abcore cannot guarantee a specific level of antibody titer in serum, as this is dependent on the antigen but will make best efforts to obtain titer. Abcore shall not be liable for its failure to perform any of its obligations resulting from circumstances beyond its reasonable control. PAYMENT Invoices should be paid no later than 30 days after the invoice date, and customers must themselves pay any bank charges that are incurred in making the payment. By signing here you agree to the Abcore Terms and Conditions. Invoicing is upon start of project, and at completion for any additional charges. If you project goes onto extension you will be invoice monthly at the end of the month services rendered. Signature: Print Name: Date: Fax Completed Pages 5-10 to or to Ship all Materials and copy of this form to: Abcore Attn: Jennifer Johnson 405 Maple Street A106 Ramona, CA Maple St., Suite A106, Ramona, CA T F P a g e 11

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