Anti-Human HER-2 (Trastuzumab) – PE
Anti-Human HER-2 (Trastuzumab) – PE
Product No.: LT1504
Product No.LT1504 Clone 4D5-8 Target HER-2/neu Product Type Biosimilar Recombinant Human Monoclonal Antibody Alternate Names ErbB-2, NEU, NGL, HER2, TKR1, CD340, MLN 19, HER-2/neu Isotype Human IgG1κ Applications FC , IHC |
Antibody DetailsProduct DetailsReactive Species Human Host Species Human Expression Host HEK-293 Cells FC Effector Activity Active Immunogen Human epidermoid carcinoma cells (A431) over-expressing EGFR. Product Concentration 0.2 mg/ml Formulation This R-phycoerythrin (R-PE) conjugate is formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.4, 1% BSA and 0.09% sodium azide as a preservative. Storage and Handling This R-phycoerythrin (R-PE) conjugate is stable when stored at 2-8°C. Do not freeze. Regulatory Status Research Use Only (RUO). Non-Therapeutic. Country of Origin USA Shipping Next Day 2-8°C Excitation Laser Blue Laser (488 nm) and/or Green Laser (532 nm)/Yellow-Green Laser (561 nm) RRIDAB_2893914 Applications and Recommended Usage? Quality Tested by Leinco FC The suggested concentration for Trastuzumab biosimilar antibody for staining cells in flow cytometry is ≤ 1.0 μg per 106 cells in a volume of 100 μl. Titration of the reagent is recommended for optimal performance for each application. Additional Applications Reported In Literature ? IHC Additional Reported Applications For Relevant Conjugates ? CyTOF® ELISA Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change. DescriptionDescriptionSpecificity This non-therapeutic biosimilar antibody uses the same variable region sequence as the therapeutic antibody Trastuzumab. Clone 4D5-8 recognizes human erbB-2. This product is for research use only. Background Trastuzumab is a monoclonal antibody targeting HER2, a 185 kDa transmembrane glycoprotein that contains an extracellular domain and intracellular tyrosine kinase activity. When it is functioning normally, the HER2 pathway supports cell growth and division. On the other hand, the over expression of HER2 propels cell growth beyond its typical range. This overexpression is associated with some cancers, namely breast and stomach, in which the HER2 protein can be expressed up to 100 times more than in typical cells. Trastuzumab induces an immune-mediated response that triggers the internalization and downregulation of HER2 making it an excellent target for immunotherapy. Several clinical studies are under way which show that anti-HER-2/neu antibodies inhibit the growth and proliferation of these tumor cells In vitro as well as In vivo. Antigen Distribution Ubiquitous expression with highest expression levels found in the kidney, skin, esophagus, and small intestine. PubMed NCBI Gene Bank ID UniProt.org Research Area Biosimilars Leinco Antibody AdvisorPowered by AI: AI is experimental and still learning how to provide the best assistance. It may occasionally generate incorrect or incomplete responses. Please do not rely solely on its recommendations when making purchasing decisions or designing experiments. Research-grade Trastuzumab biosimilars are commonly used as calibration standards or reference controls in pharmacokinetic (PK) bridging ELISA assays by serving as materials of known concentration to construct standard curves, ensuring accurate and specific quantification of the drug in serum samples. Context and Supporting Details:
Additional Information:
In summary, Trastuzumab biosimilar standards provide the critical quantitative reference needed for PK ELISA assays, enabling precise, specific, and bioanalytically comparable measurement of drug levels in serum for both clinical and regulatory studies. Flow cytometry protocols using conjugated trastuzumab biosimilars have become essential tools for validating HER2/neu expression levels and binding capacity in both research and clinical applications. These protocols leverage the specific binding properties of trastuzumab to quantitatively assess HER2 receptor density and functionality. Standard Protocol for HER2 Expression AnalysisThe fundamental protocol begins with cell preparation where target cells (commonly SKBR3, BT474, or other HER2-positive cell lines) are harvested and counted to achieve a concentration of approximately 1×10^6 cells/mL. Cells are typically resuspended in flow cytometry buffer containing PBS with 2% FBS and 0.1% sodium azide. For the binding assay, cells are incubated with conjugated trastuzumab biosimilar in serial dilutions. A standard concentration range spans from 20 µg/mL to 0.04 µg/mL using two-fold serial dilutions. The incubation is performed on ice for 30-60 minutes to prevent internalization and maintain optimal binding conditions. Detection and Analysis ParametersAPC-conjugated trastuzumab biosimilars are analyzed using red laser excitation at 650 nm, while PE-conjugated variants utilize standard PE detection channels. The mean fluorescence intensity (MFI) serves as the primary readout parameter for quantifying HER2 expression levels and binding capacity. Data analysis involves plotting 4-parameter logistic (4-PL) sigmoid curves to assess binding characteristics, with acceptable R² fitting values and low coefficient of variation (%CV) indicating robust assay performance. Relative potency calculations compare biosimilar performance against reference standards, with acceptable ranges typically falling between 80-125% as per regulatory guidelines. Quality Control and ValidationIsotype controls using humanized IgG1 kappa antibodies at equivalent concentrations ensure specificity of binding. Negative controls include cells known to have low or absent HER2 expression, while positive controls utilize established HER2-overexpressing cell lines. The protocol incorporates statistical analysis using ANOVA with Dunnett's multiple comparisons test to evaluate significant differences between biosimilars and reference standards. Parallel line analysis (PLA) provides quantitative assessment of relative potencies for regulatory compliance. Applications Beyond Expression AnalysisThese protocols extend to functional assays including antibody-dependent cellular cytotoxicity (ADCC) studies, where flow cytometry measures target cell lysis in the presence of effector cells. Additionally, the protocols support cross-presentation studies examining how trastuzumab affects antigen processing by antigen-presenting cells. The standardized nature of these protocols makes them suitable for routine quality control testing of trastuzumab biosimilars, providing cost-effective and robust methods for bioactivity assessment compared to alternative techniques like ELISA or reporter gene assays. Biopharma companies confirm the structural and functional similarity of a proposed biosimilar to its originator drug by performing an extensive suite of analytical assays, encompassing both structural (physicochemical) and functional characterization, as mandated by regulatory guidelines. Core Analytical Assays for Biosimilarity:
Analytical Similarity Assessment Approach:
Use of Leinco Biosimilars in These Studies:
Summary Table: Typical Analytical Assays for Biosimilar Comparability
Biopharma companies use this wide-ranging analytical toolbox, guided by regulatory and scientific best practice, to ensure biosimilar products are highly similar to originators, both structurally and functionally. References & Citations1. Fendly, B. et al. (1990) Cancer Research 50: 1550-1558. 2. McBride, H. et al. (2019) Pharm Res. 36(12): 177. 3. Zielinski, C. et al. (1997) Int. J. Cancer 73: 875–879 4. Valone, FH. et al. (1995) J. Clin. Oncology 13 (9): 2281-92. 5. Hynes, NE. et al. (1993) Br J Cancer. 68(6): 1140–1145. Technical ProtocolsCertificate of Analysis |
Formats Available
Prod No. | Description |
---|---|
LT1500 | |
LT1508 | |
LT1503 | |
LT1504 | |
LT1502 | |
LT1501 | |
LT1511 | |
LT1506 | |
LT1505 | |
LT1507 |
