E.coli HCP ELISA kit _ 36712ES yeasen
$645.00
In stock
SKU
36712ES96
Product Description
The use of E. coli for recombinant expression in the production of some proteins and plasmid DNA is a relatively simple and cost-effective way. The processes that use E. coli for product production are likely to contaminate host cell proteins (HCPs) from E. coli, and this contamination may reduce the efficacy of therapeutic drugs and lead to undesirable toxic or immune responses, so that the level of this contaminant needs to be minimized during purification of therapeutic drugs for use in humans or animals. ELISA is a simple to use, sensitive and objective test for HCPs. This kit uses the principle of double-antibody sandwich enzyme-linked immunosorbent assay (sandwich ELISA) to detect the residual E.coli HCP in samples . First, add the E.coli HCP standard and test sample to the Anti-E.coli HCP coated microtiter strips (36712-A) then add the diluted biotin labeled Anti-E.coli HCP antibod
y (36712-C) and finally add Streptavidin-H RP (36712-D) to form an antibody + antigen + antibody-Biotin + SA-HRP complex. Subsequently, add TMB substra te (36712-G) i n to the complex to observe color reaction after washing the complex. TMB was converted from colorless to blue and finally to yellow in the presence of termination solution (36712-H) catalyzed by HRPase. Shades of yellow were positively correlated with the amount of E. coli HCP detected in the sample. This kit can be used for the optimization of the purification process of biologics, the control of impurities in intermediate processes, and the release testing of final products. The detection range of this kit is 3.125~200 ng/mL; the lower detection limit is 2.22 n g/mL.
Features
- Highly sensitive: Detecting as little as
- 2.22
- ng/mL of
- E.coli HCP
- residuals in in-process and final biological samples.
- Ensures accuracy: The
- E.coli HCP
- residuals can be accurately detected with a recovery of
- 75
- %-1
- 25
- %.
- Specificity: Specific detection of E.coli HCP without interference from other exogenous proteins
- Stable: The difference between lot-to-lot is low, the kit property is not impacted under 37℃ for
- 7
- days.
- Easy signal collection: Using Biotin-Streptavidin system, the signal can be stably enlarged.
Applications
- Residual E.coli HCP test in biological products
Specifications
- Sensitivity
- 2.22 ng/mL (range 3.125~200 ng/mL)
- Assay Time
- <4 hours
- Assay Principle
- Two-site immunoenzymetric assay
- Signal Amplification
- Biotin-Streptavidin system
- Detection Wavelength
- 450nm-630nm
- Product Performance
- Product Performance Validation Program
- Reference standards or requirements
- YEASEN
- 1.
- Standards for the kit
- Homemade standards are available
- E.coli HCP Lyophilized powder
- 2.
- L
- inear
- R
- ange
- Assay Range
- Refer to the actual situation
- 3
- .125~2
- 00
- ng/mL
- R
- 2
- ≥0.98
- ≥0.99 and above
- Detection value CV for each concentration
- CV≤15%
- CV≤5%
- 3. Accuracy
- Recovery Rate
- 50%~150%
- 70%~130%
- and above
- 4. Precision
- Repeatability
- CV≤15%
- ＜
- 10%
- Intermediate precision
- CV≤15%
- ＜
- 15%
- 5. Exclusivity
- No interference with enzyme
- and Protein
- products from other sources
- No cross-reactivity with 9 common enzyme and Protein products
- 6. Sensitivity
- Limit of quantification
- Reliable quantification of the smallest concentrations of target substances
- 3.125 ng/mL
- Limit of detection
- Refer to the actual situation
- 2.22 ng/mL
- 7. Durability
- Reagent Stability
- Evaluating the effect of different storage temperatures on reagent performance
- 4°C for 7 days and 37°C for 7 days, with differences from T0 in the range of 70-130%.
- Instrument Compatibility
- Kits are used on different devices without affecting the test results
- Compatible with many types of Microplate Reader (eg. Molecular Devices Microplate Readers)
Components
- Components No.
- Name
- 36712ES48
- 36712ES96
- 36712
- -A
- Anti-E.coli coated microtiter strips
- 48 T
- 96 T
- 36712
- -B
- E. coli HCP Standards
- 1 vial
- 2 vial
- 36712
- -C
- Detection Antibody：Biotin-conjugated Antibodies
- 60 μL
- 120 μL
- 36712
- -D
- Streptavidin-HRP
- 20 μL
- 40 μL
- 36712
- -E
- Dilution Buffer（5×）
- 25 mL
- 50 mL
- 36712
- -F
- Wash Buffer Concentrate（20×）
- 25 mL
- 50 mL
- 36712-
- G
- TMB Substrate
- 8 mL
- 15 mL
- 36712
- -H
- Stop Solution
- 5 mL
- 10 mL
- 36712
- -I
- Plate Sealer
- 3 each
- 5 each
- The Standard (36712-B) is Lyophilized powder.
Storage
This product should be stored at 2°C ~8°C. Unopened product is valid for one year. * Upon receipt of the kit, please check whether all components are complete and immediately store them in corresponding condition. F igures Specificity Demonstration
Figure 1. The specificity of E.coli HCP ELISA kit was shown in the figure above 9 samples were estimated for the non-specific reaction with the antibodies in the kit. The concentration of the 9 samples were all below the Limit of Quantitation (LOQ) (2.22 ng/mL) Precision Display A) Repeatability Spiked sample Intra-Assay Duplicates Average Conc. (ng/mL) CV (%) S- 100 ng/mL 8 103.183 2.4% S- 25 ng/mL 8 25.677 5.7% S- 3.125 ng/mL 8 3.317 10.0% B) Intermediate precision Table 1. The precision of E.coli HCP ELISA was assayed as the above scheme Spiked sample Inter-Assay Duplicates Average Conc. (ng/mL) CV (%) S- 100 ng/mL 24 106.976 4.7% S- 25 ng/mL 24 25.470 7.2% S- 3.125 ng/mL 24 3.703 13.5% 8 duplicated were tested at 3 concentration s on the standard curve, and all the CV% were lower than 10 % ( Table 1 A). 3 lot of E.coli HCP ELISA kits were tested and the CV% were lower than 1 5 % ( Table
1B ). Accuracy Validation Three concentrations of E.coli HCP standards, high, medium and low, were spiked to 10 μg/mL of E.coli expressed purified protein (several residue-free HCP) in equal proportions, while the sample spiked at the high value was made a multiplicative dilution. HCP in the doubly diluted samples, as well as in samples of different spiked concentrations, and in unspiked samples was detected with linear and spiked recoveries ranging from 80% to 120%. Spiked recovery = measured value / (spiked standard x 50% + spiked specimen value x 50%) Table 2. Accuracy Validation Sample dilution ratio Measured value (ng/mL) Dilution linearity Sample spiking Measured value (ng/mL) Recovery rate 1×( S) 93.995 S 2.213 2 × 40.760 86.7% S+ 200 ng/mL 93.995 93.0% 4 × 19.018 93.3% S+ 100 ng/mL 51.375 100.5% 8 × 11.052 116.2% S+ 25 ng/mL 11.500 84.5% 16 × 5.190 93.9% The linear and spiked recoveries of
E.coli HCP in the doubly diluted samples, as well as the samples with different spiked concentrations, and in the unspiked samples, ranged from 80% to 12 5 % ( Table 2 ).
FAQ
- Q: What is the difference between product number 36712ES and product number 36721ES?
- A: The 36712ES item of the E. coli HCP kit is mainly used for detecting the recombinant proteins expressed by E. coli strains. It is targeted at the two E. coli strains, BL21 and Rosseta.
- The 36721ES model E.coli HCP kit is mainly used for detecting the plasmid DNA samples expressed by E.coli strains. It is targeted at strains such as DH5a, TOP10, Stbl2, and Stbl3.
Safety Data Sheet
- 36712_MSDS_HB20250310_EN.PDF
Manuals
- 36712_Manual_Ver.EN20250310.pdf
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