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Site Specific Antibody Production

$4,500.00$14,500.00

Antibody: Polyclonal and Monoclonal Antibody

Concentration: 0.25 mg/ml of purified IgG

Application: In vitro and In vivo

Reactivity: Human, Mouse, Rat

BiCell Scientific Inc has developed site-specific recognition antibodies with novel short-peptide antigen technique and hybridoma sorting technique.

Site-specific changes in protein vital for diseases are hard to detect with existing methods, such as point mutations in cancer proteins or Alzheimer’s proteins underlying disease progression. Point modifications in proteins, such as phosphorylation, acetylation, etc are often overlooked in disease diagnostics due to the lack of reliable antibodies.

BiCell Scientific Inc researchers have discovered that short-peptide antigens (13-19 amino acids) harboring single site mutation or modification allow developing polyclonal and monoclonal antibodies with specific binding to the mutated or modified site in a target protein. BiCell Scientific Inc has introduced the depletion chromatography technique with affinity column conjugated to a peptide carrying the same sequence as the antigen peptide, but not the mutation or modification, to remove unwanted bindings of antibody as an additional measure to ensure site specificity.

Protein modifications that can be targeted for site specific polyclonal or monoclonal antibody production include:
– Acetylation;   Cleavage sites;   Drug binding Isoforms;   Glycosylation;   Ligand binding;   Myristolation;
– Phosphorylation;   Prenylation;   Splice variants;   Sumoylation;   Ubiquitination;   Mutations/Polymorphisms.

Polyclonal Antibody Workflow Chart

Steps Timeline (weeks) Deliverables/Internal testing milestone QC standard
Phase I: Antigen preparation

(peptide antigen,13-19 aa length, will be designed against target protein and its modified site, synthesized and conjugated to KLH)

2 weeks / COA

 

 

 

 

Phase II: Animal immunization

(3 Sprague Dawley rats or 5 Swiss Webster mice or 2 New Zealand rabbits will be immunized with antigen; conventional protocol – three injections)

 

 

8-10 weeks 100 ul antiserum ELISA after 3rd injection
Phase III: Antiserum collection

(antiserum will be collected from one best responder animal according to ELISA report)

 

 

1-2 weeks 1 ml to 50 ml antiserum ELISA from production bleed
Phase IV: Affinity Chromatography and Depletion Chromatography

(antiserum will be depleted against affinity column conjugated with a peptide carrying the same sequence as the antigen peptide but not the modification; antiserum will then be purified with affinity column conjugated with the antigen peptide)

 

1-2 weeks 1 mg of purified polyclonal IgG IF/IHC and Western Blot

Monoclonal Antibody Workflow Chart

Steps Timeline (weeks) Deliverables/Internal testing milestone QC standard
Phase I: Antigen preparation

(peptide antigen,13-19 aa length, will be designed against target protein and its modified site, synthesized and conjugated to KLH)

2 weeks / COA

 

 

 

 

Phase II: Animal immunization

(3 Sprague Dawley rats or 5 Swiss Webster mice or 2 New Zealand rabbits will be immunized with antigen; conventional protocol – three injections)

 

 

8-10 weeks 100 ul anti-serum ELISA after 3rd injection
Phase III: Cell fusion and screening

(fusion of mouse myeloma cells with splenocytes from one best responder animal to generate hybridoma and perform antigen based hybridoma selection, enriching, and plating)

 

 

4-6 weeks Supernatant from 96 wells ELISA from supernatant
Phase IV: Expansion and antibody purification

(customer choosing one clone; expanding clone and purifying antibodies with affinity chromatography)

 

6-8 weeks One hybridoma cell line (to customer); purified monoclonal antibody from the selected hybridoma cell line (1mg)

(BiCell Scientific will bank all remaining clones; customer can request additional clones by paying a service fee for hybridoma recovery)

ELISA against antigen peptide and a peptide carrying the same sequence as the antigen peptide but not the modification

 


For Research Use Only. Not for use in clinical diagnostics.

Additional information

Antibody Deliverable

1 mg of Site-specific Polyclonal Antibody Production, 1 mg of Site-specific Monoclonal Antibody Production

Host Species

Rat, Mouse, Rabbit

Reviews

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Be the first to review “Site Specific Antibody Production”

"I am really impressed with your approach. We tried multiple times previously to create monoclonal and polyclonal antibodies to claudin-2 and MLCK1. We have had limited success generating polyclonals and no success generating monoclonals. You have generated outstanding monoclonals to both. I look forward to continuing to work with you."

Jerrold R. Turner, M.D., Ph.D.

Brigham and Women’s Hospital | Harvard Medical School

"The polyclonal antibody you generated for KIAA0408 is stunning! KIAA0408 is a novel cilium molecule that has never been studied. So, clearly there will be a lot of demand for it as we have discovered a very interesting finding and the story will be published in a high impact journal. I am strongly inclined to generate monoclonal antibody for this protein too and we should think about patenting it."

Univ.-Prof. Jay Gopalakrishnan PhD

Heinrich-Heine-Universität | Universitätsklinikum Düsseldorf

"Your ARL13B antibody works beautifully!!! We’re so happy to have a cilia-specific antibody made in rat! I can send you high resolution images to be posted on your website."

Julie Craft Van De Weghe, PhD

School of Medicine | University of Washington

"The assay is a homophilic interaction mediated cell adhesion on purified protein (in this case, immobilized purified Pcdhga9 to Pcdhga9 expressed on cell surface). Compared to control, cell adhesion is reduced in the presence of Pcdhga9 monoclonal antibody supernatants!"

Divyesh Joshi, PhD

School of Medicine | Yale University

Contact us for questions or custom requests!