ICT offers monoclonal and polyclonal antibody development, production, purification and conjugation services (under GMP conditions, if necessary). Antibodies can be developed and produced in many species including: mice; goats; chickens; rabbits; and sheep.  Antibody production can also proceed in cell culture and hollow fiber systems.  Antibody development generally takes 2 to 6 months, costs $4,000 to $30,000, and requires 10 - 170mg of antigen.

Prices and Timelines
     These sample proposals are meant to answer basic questions, and to act as project guidelines.  Depending on your specific project, the amount of antigen required may vary, conjugations may be necessary, several fusions may have to be done, the antibody may be purified, and the length of time for the project will vary.  To answer some of your questions, or to prepare a specific quote for your project, please review the antibody development questionnaire, and then contact us.  All prices are in US dollars.

#1 MoAb Devp. in 4 Mice, Production in Cell Culture
#2 MoAb Devp. in 4 Mice, GMP, Production in Hollow Fibers
#3 PoAb Devp. & Production in 2 Rabbits, GMP
#4 PoAb Devp. in 3 Rabbits, Protein A Purification, GMP
#5 PoAb Devp. & Production in 6 Rabbits with AP, GMP
#6 PoAb Development and Production in 4 Goats (GMP Compliant)
#7 PoAb Development, Purification, and Production in 3 Chickens

What are antibodies?
     Antibodies are glycoprotein immunoglobulins produced by lymphoid cells in animals in response to an antigen stimulus. Antibodies are found in serum and tissue fluids and will bind to the antigen whenever it is encountered. Antibodies are capable of binding with high specificity and affinity to the target molecule (antigen), and are stable for years when frozen at -80° C. These characteristics make them well suited in many areas, such as: immunoassays; purification protocols; flow cytometry; and immunohistochemistry.
     Monoclonal antibodies are a homogenous population of a specific antibody with one paratope. They have defined specificity and affinity toward one epitope on the antigen, are derived from one cell clone, and usually can be produced indefinitely. Polyclonal antibodies constitute a heterogeneous population of antibodies with differing paratopes for an antigen. They are commonly produced in rabbits, goats, chickens, and many other animals.
     Affinity purification of antibodies (and antigens) enhances their reactivity and utility for improved results in many applications.  The specificity and reactivity of these antibodies are critical when developing a specific assay.  ICT can provide a consistent source of reliable antibodies to add control to the antibody development, selection, and production process.
     When developing the monoclonal and polyclonal antibodies, some large molecular weight target proteins can be injected directly into the animals, and will therefore not need further modification to stimulate an immune response (i.e., conjugation to a carrier protein, see Conjugation Services ). Small molecules may require conjugation to a carrier protein to generate an adequate immune response in the host animal. 

How much does it cost to develop an antibody?
     ICT charges between $4,000 - $30,000 to develop monoclonal and polyclonal antibodies. This includes purification of the analyte and conjugation to carrier proteins (if necessary). This will produce approximately 1gm of monoclonal.  The amount of specific polyclonal antibody usually runs between 1%-15% of total serum IgG depending upon the animal's response.  All antibodies made become property of the customer. 
     It takes about 6 months to generate antibodies and requires 5 - 20 mg of purified analyte to generate monoclonal antibodies and 20 - 70 mg for polyclonal antibodies. Purified antigen is also necessary (50 - 100mg) to construct affinity columns for antibody purification.  If there is not enough purified analyte to make antibodies, ICT can produce recombinant material in E. coli or Baculovirus (see Cloning & Expression services ), try to source the molecule, or include protein purification services in a formal quote.  

Antibody Development Questionnaire

     In order to serve you better, please review and answer as many of the following questions as best you can.  Please print out the questions and fax us your answers at 952-888-8988 or simply give us a call at 1-800-829-3194 and we can go through it together.

1. If you want polyclonal antibodies:
   1. From what species (rabbit, goat, etc.)?
   2. Do you want it purified by Protein G?
2. If you want monoclonal antibodies:
   1. From what species (we assume mouse.)?
   2. Do you want it purified by Protein A?
3. n what format do you want the antibodies (ascites, cell culture supernatant, whole serum, lyophilized, etc.)?
4. Do you want it affinity purified, or purified any other way?
5. How much antibody do you need (in mg)?
6. Do you require GMP compliance?
7. What is the specific antigen that you want to make antibodies against?
8. What are the characteristics of the antigen?
   1. Is the analyte an infectious biological agent (virus, bacteria)?
   2. Are there any general toxicity issues associated with the handling or use of the analyte as an immunogen in animals?
   3. Amount of purified antigen available?
   4. Molecular weight?
   5. If the antigen is a small compound or peptide, what is the structure or amino acid sequence?
   6. Isoelectric focusing point (pI)?
   7. Extinction coefficient?
   8. Is the antigen acid labile?
   9. Storage buffer (including pH)?
   10. Are there antigen solubility issues that require the addition of guanidine or imidazole to the antigen solution (these components are toxic at high concentrations when injected IP)?
   11. Storage conditions?
   12. If the antigen is lyophilized, what stabilizers were used?
   13. Will it have to be conjugated to a carrier protein?
   14. Are there any cross-reacting or interfering substances in the buffer that may affect the generation of antibody (such as imidazole)?
9. Would you like ICT to screen the bleeds for positives?
10. Is there any other information that may affect the antibodies?

Monoclonal Antibody Sample Proposals
2 sample proposals are provided for MoAb development in 4 mice:
     Non-GMP Development and Production in Cell Culture
     GMP Development and Production in a Hollow Fiber System

     In general, it may cost between $4,000 - $30,000 to develop and produce monoclonal antibodies. This can include purification of the analyte and conjugation to carrier proteins (if necessary). This will produce approximately 1gm of monoclonal.  It takes about 6 months to generate monoclonal antibodies from the time the purified antigen is obtained and immunizations have begun.  Between 5 - 20 mg of purified analyte is required for this process.

1. MoAb Development in 4 Mice
     Development and production of monoclonal antibodies may require 2.5 mg of purified antigen per mouse (10 mg total for 4 mice). If the target antigen is of high molecular weight, it will probably not need further modification to stimulate an immune response, and can be injected directly into the mice with the proper adjuvant. If the antigen has a small molecular weight, it may have to be conjugated to a carrier protein before immunization (see Conjugation Services ).  If desired, we can quote a price for Cloning & Expression of the antigen if you do not have enough available, or it is not of sufficient purity.
     MoAb production will start after Week 21, using a cell culture system (see Section 2 below). If available, all positive clones can be tested for cross-reactivity with related compounds. This will ensure the selection of a specific clone. More than 1 fusion may be needed to develop the proper specific monoclonal antibody.  If so, an additional charge will be added per additional fusion.  Production will continue as antibody is needed. 

Week 1 First Immunization of 4 mice
Week 2 Second Immunization
Week 4 Test Bleed
Weeks 5-6 Third Immunization
Week 8 Fourth Immunization
Week 9 Test Bleed
Week 10 Cell Fusion
Weeks 12-14 Screening
Weeks 15-16 Subcloning
Weeks 17-18 Screening
Weeks 20-21 Cryopreservation of 3 Best Clones

Development (Non-GMP Compliance)...............................................$*
* If additional fusions are required, an additional charge will be added per fusion.
2. Monoclonal Antibody Production in Cell culture  
     Once the MoAb has been developed (after Week 21, see Section 1 above), production will begin using a cell culture system. This system may yield between 0.5 – 1.0 gm antibody per run.
Production in Cell culture - Research Scale, 1 run..............................$
3. Monoclonal Antibody Purification
Protein A purification of MoAb IgG, Up to 100 mL supernatant, after concentration.............................................................$*
*Additional charges are added for each additional 10 mL.
Total, Non-GMP Development and Production in Cell Culture ...................................................................... range of $8,000 - $20,000

#2 Monoclonal Antibody (MoAb) Development in 4 Mice
   GMP-Compliance with Production in a Hollow Fiber System

1. MoAb Development in 4 Mice
     Development and production of monoclonal antibodies may require 2.5 mg of purified antigen per mouse (10.0 mg total for 4 mice). If the target antigen is of high molecular weight, it will probably not need further modification to stimulate an immune response, and can be injected directly into the mice with the proper adjuvant. If the antigen has a small molecular weight, it may have to be conjugated to a carrier protein before immunization (see Conjugation Services ).  If desired, we can quote a price for Cloning & Expression of the antigen if you do not have enough available, or it is not of sufficient purity.
     MoAb production will start after Week 21, using either cell culture or a hollow fiber system (see Section 2 below). If available, all positive clones can be tested for cross-reactivity with related compounds. This will ensure the selection of a specific clone. More than 1 fusion may be needed to develop the proper specific monoclonal antibody, if so, an additional charge will be added per additional fusion.  Production will continue as antibody is needed. 

Week 1 First Immunization of 4 mice
Week 2 Second Immunization
Week 4 Test Bleed
Weeks 5-6 Third Immunization
Week 8 Fourth Immunization
Week 9 Test Bleed
Week 10 Cell Fusion
Weeks 12-14 Screening
Weeks 15-16 Subcloning
Weeks 17-18 Screening
Weeks 20-21 Cryopreservation of 3 Best Clones

Development GMP Compliance……................................................$*
* If additional fusions are required, an additional charge will be added per fusion.
2. Monoclonal Antibody Production Once the MoAb has been developed (after Week 21, see Section 1 above), production will begin using a hollow-fiber system. This system may yield between 0.5 – 1.0 gm antibody per run.
Production GMP Compliance: Hollow fiber - Research Scale, 1 run….......$
3. Monoclonal Antibody Purification Protein A purification of MoAb IgG
Up to 100 mL supernatant, after concentration......................................$*
*Additional charges will be added for each additional 10 mL
Total GMP Development and Production in a Hollow Fiber System  ...............................................................…  range of $20,000 - $30,000

Polyclonal Antibody (PoAb) Sample Proposals
     For some projects, we recommend using rabbits as the host animal in which to generate polyclonal antibodies. Rabbits tend to produce a high quality antisera from which the protein specific IgG can be easily purified, and are less expensive to maintain than larger animals, such as goats.  All rabbit polyclonal projects are done under GMP compliance.
     In general, it may cost between $4,000 - $15,000 to develop polyclonal antibodies in rabbits (which includes purification of the analyte and conjugation to carrier proteins). The amount of specific antibody usually runs between 1%-15% of total serum IgG depending upon the rabbit response.
     It takes 2-6 months to generate antibodies and requires 20 - 70 mg of purified analyte for polyclonal antibodies. Purified antigen may also be needed (50 - 100mg) to construct affinity columns for antibody purification, if necessary (see Antibody Purification ).  3 sample proposals are provided for PoAb development, production, and purification in rabbits:

#3 PoAb Development & Production in 2 Rabbits, GMP
#4 PoAb Devp. in 3 Rabbits, Protein A Purification, GMP
#5 PoAb Dev. & Production in 6 Rabbits with Affinity Purification, GMP

#3 PoAb Development & Production in 2 Rabbits, GMP

1.  Animal Purchase (2 rabbits)………………………................…$
2. PoAb Development in 2 Rabbits   Development and production of polyclonal antibodies in rabbits may require 6 mg of purified antigen per rabbit (12 mg total for 2 rabbits). After Week 6, immunizations and test bleeds will continue until antibody titers reach levels necessary for production. Immunizations will be made once a month, and test (or production) bleeds taken 2 weeks following the immunizations. Rabbit polyclonal development and production normally takes 10-14 weeks, with production bleeds generating 15-25 mL of serum per rabbit per bleed.

Project Initiation Pre-immunization Test Bleed and First Immunization
Week 1 Second Immunization
Week 2 Third Immunization
Week 4 Fourth Immunization
Week 6 Test Bleed and Titration of Test Bleed
Week 8 Immunization
Week 10 Test Bleed and Titration of Test Bleed
Week 12 Immunization
Week 14 Production Bleed

Polyclonal Development (2 rabbits over 4 months)……………………........$

3. PoAb Production  Once antibody titers reach the desired level, the on-going immunization process will require an additional 10 mg of antigen (for 2 rabbits). Injections will be given monthly, and production bleeds will be collected 2 weeks later. A minimum of 4 months of production bleeds are recommended. The serum can be aliquoted and stored at –80c indefinitely and purified as antibody is needed.
PoAb Production (2 rabbits, 4 months).............................…...........…..$
Total PoAb Production..............................................$4,000 - $6,000

#4 PoAb Development and Production in 3 Rabbits
with Protein A Purification, GMP

1.  Animal Purchase (3 rabbits)……………………………...………...…$
2. PoAb Development in 3 Rabbits  Development and production of polyclonal antibodies in rabbits may require 6 mg of purified antigen per rabbit (18 mg total for 3 rabbits). Starting after Week 8, immunization boosts are given once a month, with production bleeds taken 2 weeks following the immunizations. Rabbit PoAb development and production normally takes 12-14 weeks, with production bleeds generating 20-25 mL of serum per rabbit per bleed.

Week 1 Pre-immunization Test Bleed and First Immunization
Week 3 Second Immunization
Week 5 Third Immunization
Week 7 Fourth Immunization
Week 8 Test Bleed and Screen
Weeks 9-14 Boost and Production

PoAb from 3 rabbits………………………………….........…..........….$
3. PoAb Production  Once antibody titers reach the desired level, the on-going immunization process will require an additional 15 mg of antigen (for 3 rabbits). Injections will be given monthly, and production bleeds will be collected 2 weeks later. A minimum of 4 months of production bleeds are recommended. The serum can be aliquoted and stored at –80c indefinitely and purified as antibody is needed.
PoAb Production (3 rabbits, 4 months).............................…...........…..$
4. Protein A Purification  Total IgG can be purified by Protein A. Minimum charge for Protein A purification (up to 350 mL of serum)……......$500*
*An additional charge will be added for each additional 1.0 mL
Total PoAb Production and Purification in 3 Rabbits.... $4,000-$9,000

#5 PoAb Development and Production in 6 Rabbits 
with Affinity Purification, GMP

1. Animal Purchase (6 rabbits recommended) …………..........…… $
2. PoAb Development (6 rabbits over 4 months) …...................…... $
The development process requires 40 mg of purified antigen for 6 rabbits (and testing). After Week 6, immunizations and test bleeds will continue until antibody titers reach levels necessary for production. Immunizations will be made once a month, and test (or production) bleeds will be taken 2 weeks after each immunization. Rabbit PoAb development and production normally takes 10-14 weeks, with production bleeds generating 15-25 mL of serum per rabbit per bleed. In some cases, production bleeds have been initiated at 10 weeks.

Project Initiation Pre-Bleed and First Immunization
Week 1 Second Immunization
Week 2 Third Immunization
Week 4 Fourth Immunization
Week 6 Test Bleed and Titration of Test Bleed
Week 8 Immunization
Week 10 Test Bleed and Titration of Test Bleed
Week 12 Immunization
Week 14 Production Bleed

3.  PoAb Production (6 rabbits, 4 months) ........................................... $
Once antibody titers reach the desired level, the on-going immunization process will require an additional 30mg of antigen (for 6 rabbits). Injections will be given monthly, and production bleeds will be collected 2 weeks later. A minimum of 4 months of production bleeds is recommended. The serum can be aliquoted and stored at -80°C indefinitely and purified as antibody is needed.
4.  Affinity Purification of PoAb from Rabbits 
Affinity purification is a 2-step process. Initially, the IgG is purified by Protein A. The specific PoAb will then be affinity purified. The amount of specific antibody usually runs between 1%-15% of total serum IgG depending upon the rabbit response. 25-100mg of purified antigen is needed for construction of an affinity column. Columns often last several years and can be reused many times, depending on antigen stability when coupled to the gel matrix. 
There is 1-time column preparation fee ($400 - $1,000) for each antigen. Cost of affinity purification is based on each mL of serum, with a minimum charge of $1,000 (up to 250 mL serum). 
Column construction (per antigen)................................. up to $1,000
Minimum affinity purification charge (up to 250 mL of serum)….$ 1,000*
*Additional charges will be added for each additional 1.0 mL
Total PoAb Cost, 6 rabbits with Affinity Purification ... $8,000-$15,000

#6 PoAb Development and Production in 4 Goats (GMP Compliant)

     If there is a high expected demand for a polyclonal, we recommend using goats as the host animal in which to generate polyclonal antibodies (rather than rabbits or chickens). Goats are large enough to produce large volumes of antibody, yet small enough to be cost efficient.  All goat polyclonals are made under GMP conditions.
     In general, it may cost between $7,000 - $30,000 to develop polyclonal antibodies in goats (which includes purification of the analyte and conjugation to carrier proteins). The amount of specific antibody usually runs between 1%-15% of total serum IgG depending upon the goat response.
     It takes 2-6 months to generate antibodies and requires 20 - 70 mg of purified analyte for polyclonal antibodies. Purified antigen may also be needed (10 - 100mg) to construct affinity columns for antibody purification, if necessary (see Antibody Purification ).

1. Animal Purchase (4 goats) ……….….......................................…$
If the analyte is of high molecular weight (greater than 20,000 daltons), it should not need further modification (i.e., polymerization, or conjugation to a carrier protein) to stimulate an immune response (see Conjugation Services). It can be injected directly into the goats with the proper adjuvant. Once the polyclonals have been produced, the specific IgG will be affinity purified. On-going production of polyclonals will continue as necessary.
2. PoAb Development (4 goats) …….........................................……$
The development process requires at least 15 mg of antigen per goat. Injections and test bleeds will continue until antibody titers reach levels necessary for production.

Project Initiation Pre-Bleed and First Injection
Week 1 Second Injection
Week 2 Third Injection
Week 4 Fourth Injection
Week 6 Test Bleed and Titration of Test Bleed

3. PoAb Production (4 goats, 6 months) ………….......................…..…$
Once the antibody titers are at the desired level, production of polyclonals requires an additional 15 mg of antigen per goat. Injections will be given monthly, with production bleeds collected 2 weeks after each injection. A minimum of 6 months of production bleeds may be recommended. The serum can be aliquoted and stored at -80°C indefinitely and purified as antibody is needed.
4. PoAb Affinity Purification……………………............................…$
Affinity purification is a 2-step process, beginning with the general purification of IgG by ammonium sulfate precipitation. Once the IgG has been isolated, the analyte-specific IgG will be affinity purified. Construction of the affinity column requires 10 - 100 mg of antigen.
Total Cost of Development, Production, and Affinity Purification of Goat Polyclonal Antibodies ………… range of $7,000 - $30,000

     For some analytes, we recommend using chickens as the host animal in which to generate polyclonal antibodies. Chickens tend to produce high-quality antibodies from which the protein-specific IgG can be easily purified from egg yolks (chicken IgG is referred to as IgY).  As the PoAb is collected from the yolk, repeated bleeds are not necessary, making it easy and cost efficient to collect antibody from the egg.  All chicken PoAb projects are done under GMP conditions.
     In general, it may cost between $4,000 - $15,000 to develop polyclonal antibodies in chickens (which includes purification of the analyte and conjugation to carrier proteins). The amount of specific IgY usually runs between 1%-10% of the total IgY protein content of the yolk, depending upon the immune response.
     It takes 3-6 months to generate a humoral antibody response high enough to merit egg collection and processing.  A typical 3 - 6 hen immunization group will require 15 - 40 mg of purified analyte for injection. Purified antigen may also be needed (10 - 100mg) to construct an affinity column for specific IgY antibody purification, if necessary (see Antibody Purification ).   

1. PoAb Development ………………………………................…….$
Development and production of polyclonal antibodies may require 6 mg of purified antigen per chicken (18 mg total for 3 chickens).  If titers are acceptable, daily egg collections will begin after Week 6 (the IgY will be purified from the yolk). Once egg collection begins, injections will be made monthly.

Week 1 Pre-immunization Test Bleed and First Immunization
Week 2 Second Immunization
Week 4 Third Immunization
Week 5 Test Bleed and Screen
Weeks 9-14 Daily egg collection

2.  PoAb Production (3 chickens, 4 months) ….................................. $
Once antibody titers reach the desired level, the on-going immunization process will require additional antigen.  Injections will be given monthly, and eggs collected thereafter.  The yolks can be processed and stored at -80°C indefinitely and purified as antibody is needed.
3. Affinity Purification of Polyclonal IgY from Chickens Eggs
Affinity purification of chicken antibody is a 2-step process.  Initially, the IgY will be purified from the egg yolks using a proprietary extraction technique. The specific PoAb will then be affinity purified.  10 - 100mg of purified antigen is needed to construct an affinity column; columns often last several years and can be reused many times, depending on antigen stability when coupled to the gel matrix. There is 1-time column preparation fee ($400 - $1,000) for each antigen. Cost of affinity purification is based on each dozen of processed egg yolks.
Initial purification of 3 dozen eggs......................................................$
Affinity Column construction (per antigen)…………….…$400 - $1,000
Affinity purification, up to 3 dozen eggs...............................…............$*
*An additional charge will be added for each additional dozen eggs…….$
Total Chicken PoAb Development, Production and Purification…....  
                                                                                   $4,000 - $15,000

Need an antibody developed, purified, or conjugated? 

Call us 800-829-3194

How do I have ICT develop an assay for me?

1. Read the background information on our website. 
2. Read and fill out our Assay Development Questionnaire as best you can.
3. Contact us to briefly discuss your project and determine how our capabilities can meet your needs ( Contact ICT , 1-800-829-3194) .
4. Request a confidentiality agreement (which we can email - fax - mail to you). Review, sign, and send it back to us, or send us your confidentiality agreement for us to review.
5. Discuss the project with us in further detail so we can prepare a reliable quote.
6. Accept the quote, pay the front fee, and we’ll get started as soon as possible!