Caprico Biotechnologies, Inc. was founded in 2013 by a group of experienced scientists with broad research experience in immunology. We saw a market need to provide a range of antibody related tools and services to support basic and clinical immunological researchers. Our vision and motivation is to become a trusted and leading solution provider to the immunotherapy industry by developing innovative products that provide adequate evaluation tools for the treatment of cancer and autoimmune diseases in academic and clinical studies.


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New products for PNH clinical research!!!

FLAER

Paroxysmal nocturnal hemoglobinuria (PNH) is a stem cell disorder caused by a mutation of the gene involved in the synthesis of the GPI (glycosylphosphatidylinositol) anchor of a group of surface proteins on circulating cells. Laboratory findings in PNH include typical findings of hemolytic anemia, loss of GPI-anchored proteins.

Flow cytometry will show reduced levels of GPI-anchored proteins on peripheral blood cells.

ASR-grade FLAER (Fluorescein-labeled Proaerolysin), along with antibodies CD45CD55CD59CD14 and CD15, can be used to detect PNH clones (FLAER-negative cells) within the monocytes and granulocytes lineages in the application of multi-color flow cytometry analysis.

The analytical combination of CBI CD55 & CD59 assays can be used for detecting PHN clones in red blood cells. Flow Cytometry labs evaluate their own verified and optimized assay set up.

We are happy to provide you with free samples for your internal evaluation!

Great Performance Panel:

FLAER iFluor™488CD59 PE, CD45 PerCP, CD55 APC

Fig. 1. The common PNH flow panel is established by using Caprico products (FLAER iFluor™488, CD59 PE, CD45 PerCP, CD55 APC)  in RBC-lysed fresh blood from healthy donors.

FLAER iFluor™488 & FLAER mFluor™450 both are available right now!

Fig.2. Flow data of FLAER iFluor™488 (Right red histogram) and FLAER mFluor™450(Left red histogram) of lymphocytes gated populations. Unstained lymphocytes are used as FMO control (blue histogram).

Related products of multi-color flow panels are also available.

*ASR: Analyte Specific Reagent. The analytical and performance characteristics of this ASR product are not established.

TRBC1

FITC

TRBC1

APC

TRBC1

PE

  • A single TRBC1 antibody, JOVI.1, assay identifies the clonality of T cell subsets with a highly sensitivity.
  • Neoplastic T-cell populations can be distinguished from nonneoplastic T-cells.

Figure 1. Comparison of clonality of reactive T cells and neoplastic T cells by TRBC1 (clones JOVI.1) FITC. Left: Reactive T cells show a polytypic pattern with 42.6% of cells positive for TRBC1 and 57.4% of cells negative for TRBC1. Right: Neoplastic T cells show a monotypic pattern with 8.7% of cells positive for TRBC1. Evaluated by Wei Wang from MD Anderson Cancer Center.

T Cell Clonality

Mini Review

TRBC1, an Addition Makes Rapid and Easy T Cell Clonality Assessment for Your Assay

T cell clonality testing has important clinical and research value. The gene rearrangement of the TCR β chain locus involves the selection of one of two mutually exclusive TRBC genes, and one of 52 TCR-Vβ genes. TCR β chain constant region is encoded by two genes, namely T-cell receptor β chain constant region 1 (TRBC1) and T-cell receptor β chain constant region 2 (TRBC2). Nonpathological polyclonal T cells express a mixture of TRBC1 and TRBC2, while malignancy T cells are usually monoclonal for one β chain constant region variant 1. Both NGS-based TCR γ PCR and Vbeta repertoire analysis are highly complex, expensive, and labor-intensive. Due to poor specificity or lack of immunophenotypic clonal characterization, it provides ambiguous information in solving clonality 2.

JOVI.1 is an antibody developed for CAR-T treatment of T-cell lymphoma, showing that the expression of TRBC1 in healthy peripheral blood samples and peripheral T-cell lymphoma is significantly different 3. Studies have also shown that 25% to 47% of peripheral blood T cells express TRBC1. Recently, several leading clinical laboratories have shown important data for clonality evaluation using JOVI.1 antibody for TRBC1 assay 4-7, proving that, compared with NGS-based TCR γ PCR or Vbeta repertoire analysis, flow cytometric TRBC1 assay can provide comparable sensitivity and specificity, but faster and lower cost. The TRBC1 test provides real-time information about the T cell clonality of T cell subsets defined by the immunophenotyping. In all tested specimens, including peripheral blood and bone marrow 4,5,7, lymph nodes7, as well as tissues and body fluids 6, the neoplastic T cell population with restricted (monotype) TRBC1 expression can be distinguished from non-neoplastic T cells.

The expression of TRBC1 in all healthy donor blood is polytypic. The ratio of TRBC1+ to TRBC1– T cells is approximately in in the range of 1:1 to 1:2 (Figure 1). According to the observations from multiple laboratories, the cut-off value of neoplasms (monotypic TRBC1 expression) on which the phenotypic distinction from background benign T-cells is >85% or <15% 5

Application:

  1. The expression of TRBC1 can only be assessed in CD3-positive, TCRaβ-positive T cells, therefore accurate gating of immunophenotypically distinct T-cell subsets is critical; CD4/CD8-double negative populations should be divided into subsets of α/β and γ/δ-expressing T cell.
  2. Flow cytometric assessment of TRBC1 expression may be used to distinguish subsets of tumors that have obvious morphological overlap with other reactive or neoplastic processes.

Limitation of TRBC1 assessment

  • Surface CD3-negative neoplasms and/or TCR γ/δ positive T cells
  • B cells or NK cells and Benign thymocytes

TRBC1 assessment provides an opportunity for a simplified immunophenotypic evaluation of T-cell clonality, which can be easily implemented in the clinical workflow of routine flow cytometry practice.

Panel and Materials:

T cell Panel with TRBC1

mFluor450 mFluor540 iFluor488/FITC PE PerCP-Cyanine5.5 PE-Cyanine7 iFluor647/APC APC-iFluor700 APC-Cyanine7
CD7 CD45 TRBC1 TCRγ/δ CD2 CD3 CD5 CD4 CD8
1030146 1016166 113315 —- 100266 105386 112146 1120176 110996

Reference:

 

  1.  Mahe, E., Pugh, T. & Kamel-Reid, S. T cell clonality assessment: past, present and future. J Clin Pathol 71, 195-200, doi:10.1136/jclinpath-2017-204761 (2018).
  2. Rosati, E. et al. Overview of methodologies for T-cell receptor repertoire analysis. BMC Biotechnol 17, 61, doi:10.1186/s12896-017-0379-9 (2017).
  3. Maciocia, P. M. et al. Targeting the T cell receptor beta-chain constant region for immunotherapy of T cell malignancies. Nat Med 23, 1416-1423, doi:10.1038/nm.4444 (2017).
  4. Novikov, N. D. et al. Utility of a Simple and Robust Flow Cytometry Assay for Rapid Clonality Testing in Mature Peripheral T-Cell Lymphomas. Am J Clin Pathol 151, 494-503, doi:10.1093/ajcp/aqy173 (2019).
  5. Shi, M. et al. T-cell clones of uncertain significance are highly prevalent and show close resemblance to T-cell large granular lymphocytic leukemia. Implications for laboratory diagnostics. Mod Pathol, doi:10.1038/s41379-020-0568-2 (2020).
  6. Berg, H. et al. Flow cytometric evaluation of TRBC1 expression in tissue specimens and body fluids is a novel and specific method for assessment of T-cell clonality and diagnosis of T-cell neoplasms. Cytometry B Clin Cytom, doi:10.1002/cyto.b.21881 (2020).
  7. Shi, M. et al. Single Antibody Detection of T-Cell Receptor alphabeta Clonality by Flow Cytometry Rapidly Identifies Mature T-Cell Neoplasms and Monotypic Small CD8-Positive Subsets of Uncertain Significance. Cytometry B Clin Cytom 98, 99-107, doi:10.1002/cyto.b.21782 (2020).

A fluorophore is a fluorescent chemical that can re-emit light upon excitation. Fluorophores can be used in conjugation with protein or oligonucleotides to generate fluorescent markers for detecting the expression of proteins and nucleic acid. Fluorescent labels functionally accept light energy of a given wavelength (e.g., from a laser) and re-emit energy at longer wavelength. These two processes are known as excitation and emission.

Different types of fluorophores are used in flow cytometry based on the instrument setting (see lasers and detection channels in Figure 1), the feature of fluorophores, and markers needed in designated panels such as four, six, or ten color panels.

 

Figure 1 Brand names of Flow Cytometer and its laser and channel setting.

 


Figure 2 Caprico Biotechnologies’ 10 Color Panel, and the Spectrum of Most Used Fluorophores.

 

Dyes such as FITC, PE, APC and PerCP have become the standard for many years. Tandem dyes, comprising a small fluorophore covalently coupled to a larger fluorophore, provide maximum utilization of specific equipment, increased detection channels for each laser, and generate more information for valuable, sometimes very limited, samples.

Commonly used fluorophores for surface or intracellular epitope detection are described in Figure 2.

There are many fluorophores that have potential applications in flow cytometry. A new generation of small molecule fluorophores such as Alexa Fluor, iFluor, and mFluor are rapidly replacing many traditional fluorophores, providing users with greater light stability and brighter fluorescence. CBI offers these high-performance fluorescent dyes: iFluor™ and mFluor™ series.

iFluor™ Fluorophores

iFluor™ fluorophores are optimized for strong fluorescence, high photostability and pH independence, and are available to span the full UV-visible-IR spectrum. iFluor™ 488 has spectral properties essentially identical to Alexa Fluor® 488. iFluor™ 488 conjugates are prepared using a highly purified single rhodamine isomer which significantly improves lot to lot consistency. iFluor™ 647 dyes are spectrally similar to Alexa Fluor® 647 and DyLight™ 650 dyes.

mFluor™ Fluorophores

mFluor™ fluorophores are developed specifically for flow cytometry applications, to enable optimal multicolor detection. These dyes show large Stokes shifts and excellent water solubility. mFluor™ dyes span the full UV-visible spectrum and are designed to maximize excitation efficiency with one of the major light sources in flow cytometers such as the violet laser (405 nm) or the blue laser (488 nm). They are excellent alternatives to the phycoprotein-based tandem dyes that are quite difficult to couple to antibodies or other biomolecules.

See comparison data click here.

Caprico manufactures RUO and ASR flow cytometry reagents. With strong expertise on bioconjugation, scientists at Caprico also provide antibody conjugation services of more than 20 different type of fluorophores, including FITC, R-PE, PerCP, APC, PerCP-Cy5.5, PE-Cy5, PE-Cy7, APC-Cy7, Biotin, mFluor450, mFluor540, iFluor488, iFluor594, iFluor647, iFluor680, iFluor700, iFluor790, PE-iFluor594, APC-iFluor700.

Caprico Biotechnologies, Inc. (CBI) is an ISO9001:2015/ISO13485:2016 certified facility providing high-quality purified and fluorochrome-conjugated anti-human antibody products used in flow cytometry applications for both clinical research (RUO) and clinical diagnostics (ASR). As a start up company located in Norcross, GA, CBI is fueled by a group of highly skilled PhD scientists. We currently offer products conjugated with 16 fluorochromes derived from greater than 100 clones, useful for immunophenotyping of normal, activated (infection), and abnormal (autoimmune diseases, cancer, etc) human immune cells through surface or intracellular staining. CBI products can also be used in pre- and post-treatment monitoring in patients with a variety of diseases such as leukemia and lymphoma, PNH, MDS, etc.

All CBI products are produced from well-known clones and their quality or staining performance is equivalent to the respective products of major competitors. Typical representative comparative flow cytometry analyses for T cells, B cells, and monocytes are shown in Figure 1.

CBI also offers flow cytometry services for academic laboratories or pharmaceutical companies.

Caprico Biotechnologies, Inc. (CBI) provides high quality ASRs for clinical flow cytometry applications. Our ASR products are manufactured under strict quality standards at each step of the manufacturing process in our cGMP facility in Atlanta GA, USA. All ASRs are tested to meet specifications and ensure consistent quality lot-to-lot.

ASRs are building blocks for Laboratory Developed Test (LDT) used for diagnostics testing in CLIA/CAP high complexity laboratories.

Caprico Biotechnologies offer custom services on the following;

  • Antibody Design and Development
  • Custom Conjugation
  • Assay Design and Development

Caprico Biotechnologies follows strict cGMP / ISO / GLP standards to provide you with the highest quality custom antibodies saving you time and money. Our custom hybridomas undergo rigorous testing to ensure the final product meets your exact demands.

Monoclonal antibody design and development is lending researchers the opportunity to create highly specific, customizable mAbs for both RUO and ASR purposes. Start with a free consultation and let us take it to the step of your choosing.

Consultation:

Choose the production quantity that best meets your needs:From Idea to Antibody

Caprico Biotechnologies offers a wide range of antibody conjugates, including the new iFluor and mFluor fluorophores in addition to more classical conjugates like HRP and Biotin. Our highly specialized conjugation team will custom conjugate in-house proteins on demand, or proteins you provide to us.

CBI produces and conjugates all of its own antibodies. Each antibody is rigorously validated specifically for flow cytometry applications in an ISO9001 / ISO 13485 certified facility. CBI conjugates FITC; PE; PerCP; APC; Biotin; PerCP-Cyanine 5.5; PE- Cyanine 7, APC-Cyanine 7 and a variety of other fluorophores. Recently, CBI has partnered with AAT Bioquest®, a producer of fluorescent labeling dyes to offer a unique high-performance solution for immunophenotyping and immunomonitoring.

iFluor™ Dyes
iFluor™ dyes span the full UV-visible-IR spectrum. iFluor™ dyes have improved labeling performance compared to the classic fluorescent labeling dyes such as FITC, TRITC, Texas Red® and cyanine dyes

mFluor™ Dyes
mFluor™ dyes span the full UV-visible spectrum and designed to be maximally excited by one of the major light sources in flow cytometers such as the violet laser at 405 nm or blue laser at 488 nm.

Caprico Biotechnologies has a team of highly talented of molecular biologists, immunologists, and chemists who use the latest technologies to design and develop custom assays to meet your needs.

CBI offers and extensive array of assay design and development services including:

  • Apoptosis Quantification
  • Enzymatic Assay Development
  • Pharmacological Profiling
  • Protocol Optimization
  • Viral Detection and Quantification
  • Cytotoxicity Assay
  • Immunotoxicity Assay

 

Let the CBI Research and Development Team design and develop molecular solutions for identifying your biomarkers of interest according to our strict adherence to cGMP / ISO / GLP standards.

Flow Cytometry

Caprico can be a part of your clinical trial/research projects

Caprico Biotechnologies, Inc. is an ISO certified cGMP facility producing high-quality fluorescent-conjugated RUO and ASR anti-human antibodies suitable for academic and clinical research projects and clinical diagnostics. We offer a range of flow cytometry services including:

  • Pre-Clinical Flow Cytometry assay development and services
  • Cellular assays including characterization of cellular response to viruses, drugs, vaccines, and other entities
  • Development of potential biomarkers for diagnostic use
  • Companion diagnostics assay development for immunotherapies, cancer treatment regimes, and immunomonitoring

Our Analytical Department is equipped with flow cytometers capable of performing analysis up to 13-color in either single sample tube or 96-well format useful for detection of any type of human cell isolated from whole blood or other bodily fluids.

What Caprico can do for you

  • Caprico will provide cGMP standards ensuring optimal care of all samples and
    providing flow cytometry analysis as needed.
  • Caprico will provide you with high quality ready-for-use immunophenotyping data
    derived from your flow cytometry analysis.
  • Caprico will help design flow panels based on your specific project goals and
    needs.
  • Caprico offers multiple immunophenotyping panels which are well defined and
    optimized based on high quality reagent, including our in-house fluorescent con
    jugated antibodies (ASR and RUO grade).
  • Caprico guarantees competitive pricing significantly reducing the cost compared
    with academic or any other competitor institutions.
  • Caprico will always provide your data on the agreed upon timeline for each
    specific project.

Our Service Portfolio includes:

  • Cell immunophenotyping including peripheral blood and adherent cancer cells
    Immunoanalytical Studies
  • Bioanalytical Studies
    Flow Cytometry, Immunophenotyping,
  • Bead Based Detection methods, Intracellular Protein Analysis,
    Cellular Functional Analysis,
  • Apoptosis – FACS (Annexin V & PI/7-AAD)
    BrdU S-Phase Analysis
    Cell Cycle Analysis
  • Bioassays: Method Development & Validation Process and product related
    Impurities
  • Stability & Release Testing
    Intracellular Cytokine Secretion (ICS)

FAQs

Q: What applications are your products valid for?

A: All antibodies directed against human CD antigens are validated for use in flow cytometry. Please view the TDS for your specific product to verify which other applications your antibody is suitable for, as well as storage conditions, QC data, and much more.

Q: What are the optimal storage conditions for your antibodies?

A: Unless noted otherwise in the TDS, all conjugated antibodies should be stored at 4°C. The TDS for the product in question will contain the information outlining the appropriate storage conditions. It should be noted that conjugated antibodies should never be frozen, if possible, unless otherwise noted by that vendor’s specifications.

Q: Are Caprico’s anti-human antibodies cross-reactive with non-human primate species?

A: Caprico Biotechnologies offers multiple antibody clones which are cross-reactive with different non-human primate (NHP) species. Species reactivity and conjugates tested can be found in the respective TDS, some data are available in the resources section.  For detailed information about our NHP reactive products and the possibility of obtaining free product in exchange for test data, please contact our technical support.

Q: What volume of antibody do I need to add to my samples to guarantee accurate data and appropriate results?

A: The recommended test volume is indicated on the outer label of the product and on the TDS for that product. We recommend that every lab carries out an initial titration study before running your samples to ensure that the optimal concentration is selected for your application.

Q: How do I select an appropriate isotype control for any one of your products?

A: The isotype of any of our antibodies can be viewed in the “Isotype” field next to the clone name. Also, on the TDS for any given antibody, the “Host/Isotype” field will inform you about the isotype of that particular antibody. Once the correct isotype has been identified, just type “Isotype Control” or similar text into the search bar on the webpage and select the appropriate isotype control with the correct format (purified, FITC, PE, PerCP, etc.)

Q: What can I do to preserve my samples if they cannot be run immediately?

A: It is always best to use fresh samples, however there are measures you can take to preserve your samples for a later date. After completing your staining protocol, incubate your samples in fresh 1% paraformaldehyde for 5 to 10 minutes at room temperature. After incubation is complete, place your samples at 4°C without light exposure for up to 3 days.


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