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Membrane Protein Extracellular Loop 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 against the extracellular loop (ECL) domains in membrane proteins with novel short-peptide antigen technique and hybridoma sorting technique.

Multi-spanning membrane proteins, such as ion channels, GPCRs, ligand receptors, cell adhesion molecules, and etc, are important targets of ~50% of all small-molecule drugs but only 3 therapeutic monoclonal antibodies are available due to the challenges of targeting the extracellular loop (ECL) domains in membrane proteins. Traditional approaches of targeting the ECL domains in membrane proteins involve immunizing animals with full-length membrane proteins assembled into liposomes. Such approaches lack the site specificity in ECL domains so often require extensive epitope mapping in order to identify the right antibody.

BiCell Scientific Inc researchers have discovered that short-peptide antigens (13-19 amino acids) can be engineered into cyclic forms resembling their endogenous structures in the ECL domains of multi-spanning membrane proteins. BiCell Scientific’s approach allows developing site-specific recognition antibodies against any sequence in the extracellular loop (ECL) domains of membrane proteins. These ECL targeting antibodies can be used as blockers, neutralizing molecules, signaling molecules, CAR-T receptors and many more, for researches and therapeutics.

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 ECL domain in membrane protein, 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 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 ECL domain in membrane protein, 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 from affinity purification

 


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

Additional information

Antibody Deliverable

1 mg of Membrane Protein ECL Polyclonal Antibody Production, 1 mg of Membrane Protein ECL Monoclonal Antibody Production

Host Species

Rat, Mouse, Rabbit

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Be the first to review “Membrane Protein Extracellular Loop 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

"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

Contact us for questions or custom requests!