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Demonstration of Formalin-Fixed Paraffin-Embedded (FFPE) Specific Antibody Production

Demonstration of Formalin-Fixed Paraffin-Embedded (FFPE) Specific Antibody Production

Left: diagram illustrating the strategy of antigen protein or epitope crosslinking and stabilizing by aldehyde. Right: IHC on mouse kidney FFPE section labeled with FFPE specific anti-claudin-16 C-terminal domain antibody.

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Formalin-Fixed Paraffin-Embedded (FFPE) Specific Antibody Production

$2,150.00 – $8,950.00

Antibody: Polyclonal and Monoclonal Antibody

Concentration: 0.25 mg/ml of purified IgG

Application: IHC on FFPE

Reactivity: Human, Mouse, Rat

BiCell Scientific Inc has developed Formalin-Fixed Paraffin-Embedded (FFPE)-specific recognition antibodies with novel cross-linking and stabilizing technique to lock the conformational state of epitope in a folded state similar to aldehyde induced fixation.

Formalin-Fixed Paraffin-Embedded (FFPE) tissue specimens are the gold standards for histology and pathology. Antibody based immunohistochemistry (IHC) labeling of FFPE tissue is a difficult technique, largely owing to the conformational alteration of antigen proteins induced by formalin.

BiCell Scientific Inc has developed a novel cross-linking and stabilizing technique to lock the conformational state of epitope in a folded state similar to aldehyde induced fixation. BiCell Scientific Inc is able to raise polyclonal and monoclonal antibodies against a cross-linked target protein or an epitope sequence. Such antibodies can dramatically improve the binding specificity and affinity of IHC on FFPE sections, which will be vital tools of molecular diagnostics for pathology and oncology.

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, synthesized, crosslinked, stabilized 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 purified with affinity column conjugated with the antigen peptide)

 

 1-2 weeks 1 mg of purified polyclonal IgG IHC on FFPE

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, synthesized, crosslinked, stabilized 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)

 IHC on FFPE

 

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

Additional information

Antibody Deliverable

1 mg of Formalin-Fixed Paraffin-Embedded (FFPE) Specific Polyclonal Antibody Production, 1 mg of Formalin-Fixed Paraffin-Embedded (FFPE) Specific Monoclonal Antibody Production
Host Species

Rat, Mouse, Rabbit

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"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

"The rabbit hybridoma supernatants of anti-APOBEC3 project are tested positive by ELISA, and we are very happy about it! We previously tried a company, Abclone. Their Project "A" has immune response that is <10,000 titer in antiserum, which would explain why there is no positive mAb after fusion. Their project "B" didn't have any immune response in rabbit."

Harshita B Gupta, PhD.

School of Medicine | UT Health San Antonio

"We have tested anti mouse T cell antiserum samples from both rabbits you sent to us.

They worked very well! Thank you!"

Victoria Gorbacheva, PhD.

School of Medicine | Cleveland Clinic

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