Host Cell Proteins Detection Kit, S. aureus system

ELISA kit for the measurement of CHO host cell proteins.

Detection kit for the determination of S. aureus host cell protein contamination bulk products. Based on the "sandwich"ELISA methods, it with quick and high sensitivity analysis.

Detect and quantify HCP concentrations at any point in your purification process;
ELISA-based: quick, familiar format with high sensitivity;
96-well plate coated with capture antibody;
HCP standards, reporting antibody, streptavidin-HRP conjugate, TMB substrate, buffers included;

Host Cell Proteins Detection

The detection kit is based off of "sandwich"ELISA methods. Host Cell Protein antibodies are immobilized to 96 well plates. Samples are added, and Host Cell Proteins are bound by the antibodies. Biotinylated Host Cell Protein antibody is then added, followed by strepavidin HRP, which binds to biotin. TMB substrate is then added, and HRP catalyzes the conversion of TMB to a colored product. The reaction is then stopped and read on a standard colorimetric plate reader.

96 reactions

TBS Wash Concentrate, 20X;
S. aureus HCP Standards Set, A-F, 1 ml;
Anti-S. aureus:HRP, 12 ml;
Stop Solution;
TMB Substrate for ELISA;
Anti-S. aureus Coated Microtiter Strips;

Bacterial

S. aureus

Staphylococcus aureus

The detection kit is based off of "sandwich"ELISA methods. Host Cell Protein antibodies are immobilized to 96 well plates. Samples are added, and Host Cell Proteins are bound by the antibodies. Biotinylated Host Cell Protein antibody is then added, followed by strepavidin HRP, which binds to biotin. TMB substrate is then added, and HRP catalyzes the conversion of TMB to a colored product. The reaction is then stopped and read on a standard colorimetric plate reader.

96-well plate

Store at 4°C.

Sodium azide will interfere with this assay and should not be used in samples or buffers.

Gel Packs

1. Use clean polypropylene test tubes or micro-centrifuge vials with caps.
2. Pipette 100µL of standards, (0-500ng/mL), controls, and samples into labeled tubes or vials as indicated on work list.
3. Pipette 200µL of anti-S.aureus HCP:HRP conjugate into each tube or vial. *These volumes of 100µL for the sample and 200µL for the conjugate are recommended,
assuming the assay is performed in duplicate. If assaying in triplicate or more, the relative volumes should be adjusted appropriately.
4. Cap, vortex, and allow to incubate for 1 hour at room temperature.
5. Transfer 125µL of the reaction mixture to duplicate coated wells in the anti-S.aureus HCP coated microtiter strips as indicated on the work list.
6. Cover or place into a zip lock plastic bag. Transfer to rotator and incubate at 400-600 rpm for 1 hour at room temperature, 24°C±4°C.
7. Dump contents of wells into waste. Blot and gently but firmly tap over absorbent paper to remove most of the residual liquid. Overly aggressive banging of the plate or use of vacuum aspiration devices in an attempt to remove all residual liquid is not necessary and may cause variable dissociation of antibody bound material resulting in lower ODs and worse precision. Fill wells generously to overflowing with diluted was solution using a squirt bottle or by pipetting in ~350µL. Dump and tap again. Repeat for a total of 4 washes. Wipe off any liquid from the bottom outside of the microtiter wells as any residue can interfere in the reading step. Do not allow wash solution to remain in wells for longer than a few seconds. Do not allow wells to dry before adding substrate.
8. Pipette 100µL of TMB substrate into all wells.
9. Place cover on plate and incubate for 30 minutes at room temperature. Do not shake!
10. Pipette 100µL of stop solution.
11. Read absorbance at 450/650nm.