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resDetect™ VH3 ELISA Kit (Residue Testing)

For research use only.

产品描述(Product Details)

Assay TypeSandwich-ELISA
AnalyteVH3 binding Protein A
Format96T
Regulatory StatusRUO
Sensitivity<50pg/mL
Standard Curve Range50 pg/mL-8100 pg/mL
Assay Time2 hr
Suitable Sample TypeFor the quantitative determination of recombinant VH3 binding protein A
Sample volume50ul

背景(Background)

Traditional protein A have affinity for both the Fc region and the Fab VH3 region of human antibodies. But VH3 interacts only with the variable heavy chain of the VH3 sequence family of the human antibody,it can be widely used in the purification of related antibody drugs.. However, during the purification , VH3 may leach from the purification column and resulte in contamination of the antibody drugs prepared. Therefore, the detection of residual VH3 in antibody drugs purified from VH3 purification column is a key quality control step in the production process of antibody drug preparations.

应用说明(Application)

The kit is developed for the detection of natural or structurally conserved recombinant forms of VH3 and a recombinant form of VH3 with very significant structural differences from natural VH3 such as MabSelect VH3 in Bioprocess manufacturing applications. It is used as a tool to aid in optimal purification process development and in routine quality control of in-process streams as well as final product .

It is for research use only.

存储(Storage)

1. Unopened kit should be stored at 2℃-8℃ upon receiving.

2. Find the expiration date on the outside packaging and do not use reagents past their expiration date.

3. The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.

组分(Materials Provided)

IDComponentsSize
RES059-C01Pre-Coated Anti-VH3 Antibody Microplate1 plate
RES059-C02Recombinant VH3 binding Protein A Standard (1μg/mL)100uL
RES059-C03Biotin-Anti-VH3 Antibody1.5mL
RES059-C04Streptavidin-HRP10ug
RES059-C0510×Sample Dilution Buffer15mL
RES059-C06Denaturation Buffer15mL
RES059-C0720×Washing Buffer30mL
RES059-C08Antibody Dilution Buffer15mL
RES059-C09Streptavidin-HRP Dilution Buffer15mL
RES059-C10Substrate Solution12mL
RES059-C11Stop Solution8mL

 

典型数据-Typical Data Please refer to DS document for the assay protocol.

VH3 TYPICAL DATA

Detection of Recombinant VH3 binding Protein A by sandwich-ELISA Assay.
Immobilized Anti-VH3 binding Protein A Antibody can bind Recombinant VH3 binding Protein A. Detection was performed using Biotin-Anti-VH3 Antibody with sensitivity of 50 pg/mL (QC tested). For each experiment, a standard curve needs to be set for each micro-plate, and the specific OD value may vary depending on different laboratories, testers, or equipments. The following example data is for reference only.

 

验证(Validation)

批内差异(Intra-Assay Statistics)

Three samples of known concentration were tested ten times on one plate to assess intra-assay precision.

VH3 INTRA-ASSAY STATISTICS

批间差异(Inter-Assay Statistics)

Three samples of known concentration were tested in three separate assays to assess inter-assay precision.

VH3 INTER-ASSAY STATISTICS

回收率(Recovery)

Add different concentrations of VH3 binding Protein A (0.2ng/mL、1ng/mL、10ng/mL) to different concentrations of Human IgG4 (Toripalimab) (20mg/mL、10mg/mL、5mg/mL), then dilute the antibodies to a reasonable range, then test and calculated the concentration of VH3 binding protein A to give the recovery rate.

VH3 RECOVERY

干扰效应(Interference effect)

We have conducted interference effect test about frequently-used buffers, they have excellent buffer compatibility. For specific buffers, it is recommended that you verify recovery to determine the minimum dilution ratio.

VH3 INTERFERENCE EFFECT

专属性(Specificity)

Host cell protein (HCP 500 ng/mL) and host cell DNA (HCD 0.5 ng/mL) of HEK293, E.coli or CHO systems were added to human IgG1 (Bevacizumab, 1mg/mL) and human IgG4 (Toripalimab, 1mg/mL), respectively, which were higher than the usual quality standard limit. Then high, medium, and low concentrations of VH3 binding Protein A were added, respectively, and the ratio of VH3 binding Protein A recovery in the VH3 binding Protein A added samples without HCP and HCD was added as the specificity verification index. verification index.

VH3 SPECIFICITY

 

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