Screening / Development Services

Screening / Development Services

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Service: ELISA and Cell-Based Assay Development (Cell Cycle, Viability)
  • Catalog Number: CRO-10012
  • Description: ELISA and Cell-Based Assay Development (Cell Cycle, Viability) ELISA and Cell-Based Assays Altogen Labs can develop a cell-based assay to quantify the effect of a client’s proprietary compound by quantitation of specific biomarkers, cytokines, or phosphoproteins in cell culture supernatant or whole-cell lysates. Altogen Labs uses a cell-based assay to analyze customer samples with a robust ELISA-based assay. The activity of the compound of interest is tested on a panel of 16 intracellular Ser/Thr kinases and 22 Tyr kinases, using phospho-specific antibodies by the determination of Substrate-specific phosphorylation activity induced by kinase activation, IC50 concentrations of Ser/Thr kinases and Tyr kinases.
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Antibody Humanization

Antibody Humanization

Creative Biomart

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  • Description:

    Creative BioLabs has extensive experience in antibody humanization. Humanization is important for reducing the immunogenicity of monoclonal antibodies derived from xenogeneic sources (commonly rodent) and for improving their activation of the human immune system. Since the development of the hybridoma technology, a large number of rodent monoclonal antibodies with specificity for antigens of therapeutic interest have been generated and characterized. Rodent antibodies are highly immunogenic in humans, which limits their clinical applications, especially when repeated administration is required. Importantly, they are rapidly removed from circulation and can cause systemic inflammatory effects as well. As a means of circumventing these problems, we have developed three antibody humanization strategies that can preserve the specificity and affinity of the antibody toward the antigen whereas significantly or completely eliminate the immunogenicity of the antibody in humans. The first approach is CDR grafting and the second approach is chain shuffling. These two methods are all based on phage display of humanized scFv variants and selection of high-affinity humanized binders through bio-panning. The third method, humanized IgG library screening, is somehow unique. We will make a library of humanized whole IgG to be displayed on the surface of mammalian cells and then high-affinity binders will be sorted by FACS.

      Features

    • CDR Grafting & SDR Grafting
    • Chain Shuffling
    • Humanized IgG library screening
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Internalizing Antibody Screening
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  • Description:

    Creative BioLabs offers high-throughput phage display library screening services for selection of receptor-dependent antibodies and peptides that can be internalized into mammalian cells. Internalizing antibodies and peptides specific for cell surface receptors, which upon binding will induce receptor-mediated endocytosis, are highly suitable vehicles for targeted delivery of drugs, toxins, enzymes or DNA into the cytosol of mammalian cells for therapeutic applications. Compared with receptor-independent antibodies and peptides that can penetrate cell membrane without binding to a receptor, receptor-mediated internalizing antibodies and peptides confer cell-specific drug delivery, the fundamental requirement for targeted therapy. Selection of receptor-mediated internalizing antibodies and peptides from phage display library can also be employed to identify cell-specific markers, the endocytosed receptors that are associated with a specific cellular function.

    We have done a large number of whole-cell based phage display library screening projects to select internalizing antibodies and peptides specific for various cell surface receptors such as EGFR, NTRK2, CTLA4, HER2, CD20, CD3, CD59, transferrin receptor and CD33. Our staff scientists also have extensive experience in discovery of novel marker receptors by selecting internalizing antibodies and peptides that are specific for certain cell types. Methodologies are developed to reduce the number of internalizing phage particles that are not target receptor-specific, to exclude phage particles that are merely bound to the cell surface and to selectively identify phage particles that are capable of crosslinking receptors, rather than merely binding.

    We found that receptor-dependent internalizing antibodies and peptides are sometimes cargo-dependent as well. An internalizing antibody/peptide that permits internalization of one cargo may not support the internalization of a different cargo. Therefore, in order to select the best internalizing antibodies or peptides for a defined cargo, a custom cargo-fusion antibody/peptide library is necessary. We employ both filamentous and T7 lambda phage systems to make cargo-fusion antibody and peptide libraries in selection of internalizing antibodies and peptides, especially for toxins, enzymes and other protein cargos.

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Membrane Protein Screening

Membrane Protein Screening

Creative Biomart

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  • Description:

    As a leading supplier for services in the biotechnology field, Creative BioMart Services for membrane protein extraction, isolation and screening are open to any project, with a focus on the demanding class of membrane proteins. Our service includes a thorough discussion and counseling of your project, so that the methods used provide optimal conditions for your protein of interest.

    Our service offering includes, but is not limited to

    • Membrane protein extraction from cell, cell lysate or cell culture supernatants.
    • Membrane protein isolation for cytoplasm membrane, mitochondrion membrane, chloroplast membrane, vacuole membrane or nucleus membrane.
    • Membrane protein screening by particles-ELISA.
    • Kinetic analysis of antibody binding by biosensor.

      We use a kind of particle to provide an alternative to living cells, membrane preparations, and detergent-solubilized proteins by offering concentrated membrane proteins in their native conformation. Particles can be used in standard membrane protein assays and are compatible with a wide variety of detection platforms.

      Creative BioMart provides technical support including experimental protocols and strict quality control using up to 16 metrics per batch. Any service package can be booked on its own, or in conjunction with others. Please contact us for more information

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Phage Display Library Screening
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  • Description:

    Our staff scientists have extensive experience in screening of phage display peptide, cDNA and scFv/Fab libraries. In particular, by conducting library screening, panning, for 4 cycles, we normally get scFv/Fab antibodies of an affinity of 10-7 By constructing serial sub-libraries of the isolated scFv/Fab antibodies, Creative Biolabs' proprietary protocol allows increase of the affinity of the scFV antibodies from 10-8 to 10-9. We have successfully obtained a scFV antibody that has an extremely high affinity of 10-12, whose binding to the antigen is essentially irreversible.

    Phage Display Libraries:

    • Libraries commercially available
    • In house premade libraries (HuScL®, HuSdL®, HuFabL®, MuScL®)
    • Custom constructed library or libraries from the customer

      Targets:

      • Purified antigens;
      • Un-purified antigens;
      • Intact cells, tissues or any in vivo selection systems required by the customer.

        Applications:

        • Identify enzyme inhibitors
        • Map epitopes on antigens;
        • Select new antibodies;
        • Discover/validate new therapeutic targets;
        • Discover interaction between ligand and receptor;
        • Select any peptide/protein binders
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Protease Substrates Screening
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  • Description:

    Phage display peptide libraries that display random 5-20 mer peptides have been widely used to determine protease substrate [sequence] specificity for a large number of enzymes and semi-purified cell extracts. This technique has been playing a critical role in designing inhibitors and activators of proteases.

    In comparison with phage display peptide libraries used in other fields, phage display peptide libraries used to identify the preferred substrate sequences of a protease usually express a fusion tag at the N-terminus of the peptide sequences, while, like in regular phage display peptide libraries, the minor coat viral protein pIII is located at the C-terminus of the peptide sequences. The N-terminal fusion tag is intended to bind the phage particles to a solid support. Scientists from Creative Biolabs have designed and built up a unique tag, HiAffi-tag, which is derived from Protein G, thus has strong binding affinity to most IgG coated on a solid surface. In contrast to His Tag and Strepavidin Binding Motifs used in such libraries, HiAffi-tag enables the best capture of the phage particles to the solid support and the least steric hindrance for proteolysis of the substrate peptides.

    During library screening against a particular protease, phage particles that express sensitive peptide sequences are cleaved off the solid support. After that, the free phages are amplified, immobilized and subjected to protease cleavage again. After several rounds of screening, selected phage clones are DNA sequenced and common features of the peptide sequences are determined. These peptide sequences are good candidates for further identification of the substrates of the protease.

    Creative Biolabs also developed proprietary in silicon screening and computer aided rational design methods to use such peptide sequences, which must fit into the active site of the protease, as leads to develop inhibitors and activators of a protease.

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Protein Ubiquitinylation Service
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  • Description:

    As a leading biotechnology services supplier, Creative BioMart offers protein ubiquitinylation services. With our expertise as a leader in the field of ubiquitinylation services, Creative BioMart has created a set of new platform technologies.

    Ubiquitin Screening Service

    Creative BioMart provides compound screening and profiling against E3 ligase cascades. Each ubiquitination cascade consists of the three enzyme components (E1 activating enzyme, E2 conjugating enzyme and E3 ligase), plus a biologically-relevant substrate. The incorporation of ubiquitin into the substrate is quantitatively detected by electrochemiluminescence, which allows for the identification of compound inhibitors and activators.

    Highlights

    • Validated assays for an expanding panel of E3 ligase cascades to screen and profile inhibitors and enhancers.
    • Cascades consist of E1, E2 and E3 enzymes and substrates.
    • Uses biologically relevant substrate.
    • Single or multiple concentration.
    • Run by the same experts who have run many years experiences.
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Stem Cell Assay Development and Screening
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  • Description:

    Creative Bioarray has a broad range of assay tools that can be implemented with PSCs or their differentiated progeny to interrogate disease-relevant biology such as cell viability, cell proliferation, apoptosis, autophagy, generation of reactive oxygen species, mitochondrial function, ion transport, pathway signaling, gene expression, protein expression and post-translational modifications.

    Our stem cell screening services enable rapid profiling of your compounds against many assay technology platforms offered by Creative Bioarray. Compounds will be profiled as 10-point dose-response curves in duplicate or triplicate.

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  • Altogen Labs
  • Creative Biomart