E.coli HCP ELISA Kit (Plasmid) _ 36721ES yeasen
$735.00
In stock
SKU
36721ES48
Product Description
The use of Escherichia coli (E.coli) for expression and amplification in the production of recombinant proteins and plasmid DNA is a relatively simple and cost-effective way of preparing biologics. However, the process of using E. coli inevitably introduces host cell protein (HCP) impurities, which can reduce the efficacy of the product and may lead to adverse toxicity or immune responses, and it is therefore necessary to ensure that the level of HCP residues in the final biologic product is as low as possible. 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 E.coli HCP residues for plasmids in samples . First, add the E.coli HCP Standard for Plasmids ( 367 21 -B ) and test sample to the Anti-E.coli HCP coated microtiter strips (36721-A) then add the diluted
biotin labeled Anti-E.coli HCP antibod y (36721-C) and finally add Streptavidin-H RP (36721-D) to form an antibody + antigen + antibody-Biotin + SA-HRP complex. Subsequently, add TMB substra te (36721-H) 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 ( 36721 I ) 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 0.5 n g/mL.
Features
- Highly sensitive: Detecting as little as
- 0.5
- ng/mL of
- E.coli HCP
- (Plasmid) residuals in in-process and final biological samples.
- Ensures accuracy: The
- E.coli HCP (Plasmid)
- 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.col
- i HCP
- (Plasmid) test in biological products
Specifications
- Sensitivity
- 0.5 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
- 3.125~200 ng/mL
- R2
- ≥0.98
- ≥0.99 and above
- Detection value CV for each concentration
- CV≤15%
- CV≤10%
- 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
- Protein
- products from other sources
- Specific detection of
- E.coli
- HCP without interference from other exogenous proteins
- 6. Sensitivity
- Limit of quantification
- Reliable quantification of the smallest concentrations of target substances
- 1.5 ng/mL
- Limit of detection
- Refer to the actual situation
- 0.5 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
- 36721ES48
- 36721ES96
- 36721
- -A
- Anti-E.coli coated microtiter strips
- 48 T
- 96 T
- 36721
- -B
- Standard: E.coli HCP (10 μg/mL)
- 50 μL
- 100 μL
- 36721
- -C
- Detection Antibody: Biotin-conjugated Antibodies (50×)
- 150 μL
- 300 μL
- 36721
- -D
- Streptavidin-HRP (100×)
- 150 μL
- 300 μL
- 36721
- -E
- Dilution Buffer 1
- 25 mL
- 50 mL
- 36721
- -F
- Wash Buffer Concentrate
- (
- 20×
- )
- 25 mL
- 50 mL
- 36721-
- G
- Dilution Buffer 2
- 15
- mL
- 30
- mL
- 36721
- -H
- TMB Substrate
- 6
- mL
- 12
- mL
- 36721
- -I
- Stop Solution
- 5 mL
- 10 mL
- 36721
- -J
- Plate Sealer
- 3 each
- 5 each
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.
Figures
Precision Display A) Repeatability Spiked sample Intra-Assay Duplicates CV （ % ） S- 100 ng/mL 10 3.24% S- 25 ng/mL 10 4.17% S- 6.25 ng/mL 10 8.26% B) Intermediate precision Table 1. The precision of E.coli HCP ELISA was assayed as the above scheme Spiked sample Inter-Assay Duplicates CV （ % ） S- 100 ng/mL 30 5.32% S- 25 ng/mL 30 9.16% S- 6.25 ng/mL 30 13.07% 10 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 A) Dilution linearity The plasmid samples amplified using E.coli cells (concentration 22.6 mg/mL, HCP concentration 55 ng/mL) were gradient diluted with sample diluent, and the HCP residue values were determined using this kit. From the following data, it can be seen that the linearity of the sample dilution determined by this kit is good, the difference between the HCP assayed value and the initial concentration after different times of dilution is small, the linear recovery is in the range of 80%~120%, and the detection value of HCP is linearly correlated with the number of times of dilution.
Table2 Sample dilution linearity Sample name HCP concentration (ng/mL) Dilution factor Detection mean (ng/mL) Actual HCP measurement (ng/mL) Recovery ratio (%) Authentic plasmid samples 55 2 22.83 45.65 83.0% 4 13.50 53.98 98.1% 8 7.35 58.81 106.9% B) Spiking Recovery Rate The plasmid samples (concentration of 22.6 mg/mL), which had been amplified using E.coli, were diluted 2-fold and mixed with equal volumes of standards of each concentration, and the spiked recoveries ranged from 80% to 120%. Spiked recovery % = (measured value of spiked samples - measured value of unspiked samples) theoretical value (amount of spiked standard) x 100%. Table3 Sample spiking recovery rate Sample Detection mean (ng/mL) Recovery ratio (%) S 23.65 S + 200 ng/mL（V:V=1:1） 117.73 94.08% S + 50 ng/mL（V:V=1:1） 48.17 98.08% S + 12.5 ng/mL（V:V=1:1） 30.75 113.60%
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 samples such as the recombinant proteins expressed by E. coli strains. It is targeted at the two E. coli strains, BL21 and Rosseta.
- The 36721ES item of the E. coli HCP kit is mainly used for detecting the plasmid DNA samples expressed by E. coli strains, and it is applicable to DH5a, TOP10, Stbl2, and Stbl3.
Safety Data Sheet
- 36721_MSDS_HB250310_EN.PDF
Manuals
- 36721_Manual_HB250310_EN.pdf
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