Services

Custom Antibody Development/Production/Generation Services

WE provide a full range of custom antibody development/production services or the life science and biotech industry.

 

Custom Polyclonal Antibody Development/Production Service in Rabbit (2 New Zealand white rabbits)

Salient Features of Custom Polyclonal Antibody Development/Production Service

  • 12 Weeks protocol to produce antibody in 2 New Zealand White Rabbits (From confirmed Pure line Parental stock).
  • This will include 7 immunizations and 3 Production bleeds per animal.
  • Pre-immune sera of 5-10 ml per animal and a total of 30-40 ml of crude immune sera per rabbit.
  • *FREE ELISA Titer at first bleed against free immunogen.

Customer will be notified of immunization schedule and tentative date of production/Progress of the Project etc.

Immunogen required for custom polyclonal antibody development/production 

  1. For Protein (Provided by customer)-3 mg, Concentration: minimum 0.5 mg/ml
  2. For peptide (Min ~5-7 mg)

IMMUNIZATION AND BLEEDING SCHEDULE

WEEKS

PROCEDURE

BLEED/RABBIT

Week 1 (Pre-bleed)
Primary Immunization

Pre-Immune Sera and Primary Immunization

~5 ml*

Week 2 (Immunize)

2nd Immunization

 

Week 3 (Immunize)

3rd Immunization

 

Week 4 (Immunize)

4th Immunization

 

Week 5 (Immunize)

5th Immunization

 

Week 6 (Bleed-1)

1st production Bleed

~10 ml*

Week 7 (Immunize)

6th Immunization

 

Week 8 (Bleed-2)

2nd production Bleed

~10 ml*

Week 9 (Immunize)

7th Immunization

 

Week 10 (Bleed-3)

3rd production Bleed

~10 ml*

 

Extention of project per month (2 bleeds/Rabbit)

Bleed-1

~10 ml*

Bleed-2

~10 ml*

 

Terminal bleed
(1 bleed per Rabbit)

Bleed-1

~20-25 ml*

 

* In our custom polyclonal development/production service we usually provide the above mentioned sera from one rabbit/project but it may vary depending upon the health status of the animal.

The above schedule of custom polyclonal antibody development/production is approximate. We can also modify it according to the customer’s need.

Additional 4 weeks is required if the custom polyclonal antibody development/production service requires peptide synthesis, mass spec, KLH conjugation. For custom polyclonal antibody development service in which the immunogen is supplied can be started immediately. 

Purification:


Protein A : 15-20 mg of purified Antibody
Peptide affinity : 2-3 mg of purified Antibody

 

 

Additional Services

  • Custom peptide synthesis
  • Recombinant protein production for custom polyclonal antibody development/production in India
  • Custom polyclonal antibody development service against Phospho/Acetylated protein in India
  • Custom polyclonal antibody development service in Mice/Rat in India
  • Validation of custom polyclonal antibody for FACS, IP, IHC and IF
  • Custom monoclonal antibody development service in India

Terms and Conditions:
 

If the immunogen is peptide then it must contain a cysteine for conjugation.

  1. If the immunogen is already conjugated or it’s in gel, kindly provide us ~1 mg of free peptide/purified protein for ELISA.
  2. Recombinant protein and purified protein must be shipped in dry ice.
  3. Kindly provide antigen in soluble form.
  4. For peptide affinity purification customer should provide additional 6 mg peptide.
  5. Biostring Inc. guarantees ELISA positive sera for the immunogen synthesized.
  6. The titre of the antisera depends on the quality of the antigen.

 

Custom Monoclonal Antibody Development/Production Service

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We are a leading custom monoclonal antibody developing company. Availability of newer reagents paves the way for development of new diagnostic assays and helps in finding potential therapies for such diseases.

 

 

A. Partial Custom Monoclonal Antibody Development/Production Service 

 

For partial custom monoclonal antibody development/production service, customer provides immunogen in the form of purified protein, fusion protein, protein extracted from bacteria, virus, gel extracted protein, recombinant protein and synthetic peptide. We require a minimum of 5 mg of the protein at a minimum concentration of 0.5 mg/ml.  

B. Complete Custom Monoclonal Antibody Development/Production Service-Peptide 

 

For custom monoclonal antibody development/production service from peptide, customer provides the sequence information of the protein (Ref. The Gene Accession number from NCBI gives details of antigen information). We go via the peptide route to produce antibody for you. This is achieved by selecting the epitopic region (possible immunogenic sites) from the sequence and immunizing the mice with the peptide conjugated to a carrier molecule. 

C. Complete Package for Custom Monoclonal Antibody Development/Production Service-Partial recombinant protein 

For custom monoclonal antibody development service from partial recombinat protein, customer provides the sequence information of the protein (Ref. The Gene Accession number from NCBI gives details of antigen information). Custom monoclonal antibody development service includes all the features of partial package for custom polyclonal service apart from the following.

1. Identification of target antigen

2. Gene Synthesis and Partial Recombinant protein purification (Upto 200 amino acid) 

There are four phases for each custom monoclonal antibody development project. Each phase has its own duration and pricing structure. Movement from one phase to the other would be discussed with the customer and depending on the progress, decision would be taken. We shall do our best to get things done faster. However, there are multiple factors those influence the progress while meeting the dead line and that has to be sorted out amicably. Customer can start a project for a particular phase with a single purchase order (PO) and move on to the next phase with a separate PO or send order including all the features at a time.

 

 

 

Phase I: Antisera Development in Mouse
Duration: 6-9 weeks 

We check the purity of the customer supplied protein in SDS-PAGE, to ensure the integrity of the protein before initiating the project.

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Choice of animal is Balb/c mice. We introduce 4 mice for each antigen. Quantity of immunogen required is minimum 5 mg. We prefer the customer to supply it in a single batch. However, it can also be accepted in two batches if the protein is difficult to express or purify. We have a standard immunization schedule specific for peptide antigen and for native protein/recombinant protein antigen. However, customer can feel free to advise us the immunization schedule depending on the antigen they supply. At the end of immunization the animals are bled and checked for the specific titer by an indirect ELISA using free immunogen as antigen and if desired, Western blot can also be done. At the end of this stage, we would send at least 20 µl of test sera sample along with the pre immune sera to the customer for testing at their end. We give guarantee of ELISA positive sera only on antigen supplied by us. For customer supplied antigen, we can attempt a few more boosters if the sera are negative or non specific in the test system, before calling it off. After results are available and acceptable to the customer, we would proceed to Phase II.

 

Phase II: Fusion
Duration: 8-10 weeks 

http://abgenex.com/images/content/Ph-2.jpgAfter customer checks the specificity and affinity of the antibody in mouse sera, we decide to go for generation of mAb using PEG fusion technology (The mouse to be fused would be specified by the customer). Splenocytes are fused with suitable myeloma partner, and hybridoma clones are raised. Growing colonies are screened for their Ab producing capacity and detected by ELISA and if desired western blot (depending on the availability of the testing materials) can also be done free of charge. Strong ELISA positive clones are selected, expanded and frozen. At this stage, a minimum of 5-10 ml of culture supernatant from positive clones is also supplied to customer for validating antibody in desirable application system. If the customer wants to wind up the project at this stage, then we supply hybridoma cells (ELISA positive) to customer. However, if the customer wants the project to be continued, we send everything at the completion of each project. Customer must be aware that we can guarantee only ELISA positive clones.

 

Phase III: Sub-cloning and isotyping
Duration: 3-5 weeks 

http://abgenex.com/images/content/Ph-3.jpgAfter thorough investigation when the validity of the antibody application is confirmed, repeated attempts would be made to generate stable single cell clone stage through limiting dilution. Sub-cloning would be continued until we get a stable clone that produces antibody continuously and specifically. At this stage, the culture supernatants from two sub-clones would be sent to customer for verification. We would also do the isotyping of the clones at this stage and preferably select IgG isotypes, unless the customer wants otherwise. We will send the live cells at room temperature to avoid any damage due to the frozen cells during dry ice shipment. We can validate the mAb for other applications; for example, FACS, IP, IHC and IF at extra charge (see Additional Services).

 

Phase-IV: Large scale production of mAb and Purification
(Min. ~2-3 mg of Purified Antibody will be supplied)
Duration: 2-4 weeks 

http://abgenex.com/images/content/Ph-4.jpgWe select the clone suggested by customer. We scale up the mAb production using spinner flasks in continuous culture method. Then we purify the mAb from culture supernatant using specialized matrix column. We provide the purified antibody along with QC data.

 

 

Recombinant Protein Production

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Contact us before starting your next custom recombinant protein production/expression/purification project, we will work with you to determine your requirements and prepare a detailed proposal.

Salient features of recombinant protein production/expression/purification service

  • ~2-5 mg of the purified protein will be supplied.
  • Q.C. of the purified protein will be carried out by SDS-PAGE and WB by anti Anti-His antibody.
  • We also produce Functional Grade Recombinant Protein.

Procedure for Recombinant protein production/expression/purification

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Stable Cell Lines Development

We provide a full range of custom services for the life science & biomedical industry in the field of drug discovery. The stably-transfected cell lines are the best available solution for a wide range of applications, such as functional studies, drug discovery assays or the production of recombinant proteins. In contrast to transient expression, stable expression allows long term, as well as defined and reproducible, expression of the gene of interest. With state of art laboratory facility and world-class expertise, we offer a comprehensive service for the development of stable cell line.

All projects begin with a project evaluation in which we discuss and review your requirements. This includes target evaluation and applications. Then we prepare a project proposal and quotation based on your needs. This will address all costs as well as the scope of work to be completed and the estimated timeline of the project. 

Our Custom Service Manager is available to consult with you at any time, and will provide you with updates in regular basis.

Procedure for development of Stable Cell Lines

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