This highly sensitive "sandwich" test kit is intended for use in the quantitative determination of antibody SN38 conjugate level in human serum or plasma. It is useful for pre-clinical and clinical pharmacology study of SN38 Antibody Drug Conjugate (ADC).
Contents of Kit
Prior to use allow all reagents to come to room temperature. Regents from different kit lot numbers should not be combined or interchanged. 1. Anti-SN38 Antibody Coated Microplate. One microplate with twelve by eight strips (96 wells total) coated with Anti-SN38 antibody. The plate is framed and sealed in a foil zipper bag with a desiccant. This reagent should be stored at 2-8°C and is stable until the expiration date on the kit box. 2. SN38 Tracer Antibody. One vial containing 12mL of ready to use SN38 Tracer Antibody in a stabilized protein matrix. This reagent should be stored at 2-8°C and is table until the expiration date on the kit box. 3. ELISA Wash Concentrate. One bottle containing 50 mL of 20-fold concentrate. Before use the contents must be diluted with 950 mL of demineralized water and mixed well. Upon dilution, this yields a working wash solution containing a surfactant in phosphate buffered saline with a non-azide, non-mercury preservative. The diluted wash solution may be stored at room temperature and is stable until the expiration date on the kit box. 4. ELISA HRP Substrate. Two bottles containing 6 mL of tetramethylbenzidine (TMB) with hydrogen peroxide. This reagent should be stored at 2 - 8°C and is stable until the expiration date on the kit box. 5. ELISA Stop Solution. One bottle containing 12 mL of stop solution. This reagent may be stored at 2 - 8°C or room temperature and is stable until the expiration date on the kit box. 6. Assay Buffer. One bottle containing 12 mL ready-to-use buffer. It should be used according to the assay procedures. This reagent should be stored at 2 - 8°C and is stable until the expiration date on the kit box. 7. Antibody Conjugated Calibrator Zero. One vial containing 30 mL calibrator zero. This reagent is used for diluting the calibrator stock to make assay calibrators, as well as for diluting test samples. This reagent should be stored at 2-8°C and is stable until the expiration date on the kit box. 8. Antibody-SN38 Conjugated Calibrator Stock-Not provided in the kit (optional). One vial containing the calibration stock of antibody SN38-conjugate which is hRS7-SN38(Sacituzumab govitecan) The DAR is 8. Refer to the vial for exact concentration of the standard. This reagent should be stored at -80°C and is stable for 12 months.
Storage
This test kit must be stored at 2°C~8°C upon receipt. For the expiration date of the kit refer to the label on the kit box. All components are stable until this expiration date.
Precision
Intra-assay Precision (Precision within an assay):3.79%. Inter-assay Precision (Precision between assays) :10.51%.
Sensitivity
The limit of quantitation(LOQ) was 10ng/mL, the concentration corresponding to the value obtained by test SN38 Antibody Drug Conjugate spiked to a low concentration of 10ng/mL under the precision CV＜15% and the recovery in the range of 80-120%. Hook: 500ng/mL.
Standard Curve
A typical absorbance data and the resulting standard curve from this SN38 ADC ELISA are represented. This curve should not be used in lieu of standard curve generated with each assay.
Citations
Publication ()
Have you cited DEIABL316 in a publication? Let us know and earn a reward for your research.
Background
Camptothecin (CPT) is an alkaloid anti-tumor drug extracted from the bark and fruits of Camptotheca acuminata. Camptothecin has good therapeutic effects on various malignant tumors such as digestive tract tumors, leukemia, and bladder cancer. Among them, SN38 is the active substance metabolized in the body of CPT-11 (a water-soluble semi-synthetic derivative of CPT).
Figure 1. Structures and action mechanisms of SN-38 based ADCs.(Source: Liu L, et al., 2021 )
Studies have found that camptothecin can specifically inhibit the activity of topoisomerase I (ToPoI). ToPoI is a ubiquitous ribozyme in the body and is overexpressed in a variety of tumor tissues. ToPoI can catalyze the relaxation of double-stranded DNA in a supercoiled state and participate in many key nuclear processes such as DNA replication, transcription, recombination and repair. process, and CPT can combine with the ToPo I-DNA cleavable complex to form a CPT-ToPo I-DNA ternary complex, which prevents DNA from being replicated and transcribed normally, and ultimately leads to the death of tumor cells. As a new specific inhibitor of topoisomerase, CPT-11 has the same main anti-tumor mechanism as CPT.
However, the extremely poor water solubility of camptothecin makes it difficult to prepare it into a suitable dosage form, and clinical studies have shown that camptothecin treatment can cause severe bone marrow suppression, diarrhea, vomiting, hematuria and other toxic side effects. These defects greatly limit its clinical application value. To this end, researchers have introduced various methods to improve its therapeutic effect and reduce toxic and side effects. Among them, SN38 ADC development is an area of great concern. Antibody-drug conjugates (ADCs) are a class of biopharmaceutical drugs designed as targeted therapies for the treatment of cancer. Unlike chemotherapy, ADCs are designed to target and kill tumor cells while sparing healthy cells. Therefore, SN38 ADC is a complex molecule composed of SN38 and a targeting antibody. It can deliver SN38 only on tumor cells through antibody-antigen specific binding. Antibodies attach to antigens on the surface of cancer cells. The biochemical reactions that occur upon attachment trigger signals in the tumor cells, which then take up or internalize the antibodies and attached cytotoxins. Once the ADC is internalized, SN38 functions to kill the cancer. In order to verify the effect of the developed SN38-ADC, it is necessary to detect the level of SN38 conjugates in serum or plasma after its use. Among them, the SN38 ADC ELISA Kit is an analytical tool used to quantitatively determine the level of SN38 ADC in human serum or plasma.
Creative Diagnostics products are for RESEARCH USE ONLY, please make sure your review is research based.
Required fields are marked with *
Terms and conditions:
We will select high-quality review customers and offer a $30 coupon for your next purchase.
All product reviews must be submitted in the English language.
Creative Diagnostics will not share any personal information of applicants, and all information will be treated with strict confidentiality and will not be sold or disclosed to a third party.
References
Structural Modification Endows Small-Molecular SN38 Derivatives with Multifaceted Functions
As a camptothecin derivative, 7-ethyl-10-hydroxycamptothecin (SN38) combats cancer by inhibiting topoisomerase I. SN38 is one of the most active compounds among camptothecin derivatives. In addition, SN38 is also a theranostic reagent due to its intrinsic fluorescence. However, the poor water solubility, high systemic toxicity and limited action against drug resistance and metastasis of tumor cells of SN38 indicates that there is great space for the structural modification of SN38. From the perspective of chemical modification, this paper summarizes the progress of SN38 in improving solubility, increasing activity, reducing toxicity and possessing multifunction and analyzes the strategies of structure modification to provide a reference for drug development based on SN38.
Antibody-drug conjugates targeting TROP-2 and incorporating SN-38: A case study of anti-TROP-2 sacituzumab govitecan
Antibody-drug conjugates (ADCs) that exploit the active metabolite SN-38, which is derived from the popular anticancer drug, irinotecan (a camptothecin that inhibits the nuclear topoisomerase I enzyme, inducing double-stranded DNA breaks during the mitotic S-phase of affected cells), represent a substantial advance in the ADC field. SN-38 has been conjugated to a humanized antibody against trophoblast cell surface antigen 2 (TROP-2), which is involved in cancer signaling pathways and has increased expression by many cancer cell types, yielding the ADC sacituzumab govitecan. By conjugating a higher number of SN-38 molecules to the immunoglobulin (drug-to-antibody ratio = 7-8:1), and giving higher (10 mg/kg) and repeated therapy cycles (Days 1 and 8 of 21-day cycles), enhanced drug uptake by the targeted cancer cells is achieved. Based on a unique conjugation method, the lactone ring of the SN-38 molecule is stabilized and the molecule is protected from glucuronidation, a process that contributes to the untoward late diarrhea experienced with irinotecan. Finally, while the ADC is internalized, the use of a moderately stable linker permits release of SN-38 in an acidic environment of the tumor cell and its microenvironment, contributing to a bystander effect on neighboring cancer cells. Here, we discuss the development of sacituzumab govitecan and clinical results obtained using it for the management of patients with advanced, refractive breast, lung, and urinary bladder cancers. Sacituzumab govitecan, which is undergoing accelerated approval review by the US Food and Drug Administration while also being studied in Phase 3 clinical studies, was granted Breakthrough Therapy status from the FDA for advanced, refractory, metastatic triple-negative breast cancer patients.
COA
Publication 
Figure 1. Structures and action mechanisms of SN-38 based ADCs.(Source: Liu L, et al., 2021 )
Loading ......
