Human IgG kit

Human IgG kit for clone selection and IgG bioprocess

Accurate & fast quantification of antibodies is critical in Bioprocess for antibody production. The HTRF® human IgG kit is designed for the quantification of human IgG in cell supernatant and the selection of the best producer clones. It offers the benefit of being much less time-consuming than conventional ELISA methods. Furthermore, this assay requires only a small amount of cell culture sample, enabling antibody screening at early stages of production.

Assay principle

This kit is a sandwich immunoassay enabling highly-sensitive quantification of human IgG , even when using a small sample volume. The assay uses two polyclonal antibodies, one (Fc specific) labelled with Cryptate and the second (FAb specific) with d2. The signal intensity  is proportional  to the number of antibody-antibody complexes formed and therefore to the human IgG concentration.

Product specifications

The simple procedure allows testing of therapeutic antibodies under high throughput, and facilitates the miniaturization process. The assay is based on a sandwich format, gives a sensitivity of 0.9 ng/mL,  and requires just one 2-hour incubation. This high sensitivity enables measurements using small sample volumes, suitable for monitoring the early stages of antibody production.

The figure on the right shows the human IgG kit calibration curve. The assay was performed in a 384-well small volume white microplate in a 20 µL total volume, using 10µL human IgG calibrator diluted in PBS 0.1 M and 5 µL of each conjugate. The two HTRF conjugates may be pre-mixed and added in a single dispensing step to further streamline the protocol. After 2h of incubation, the signal was read on a PHERAstarFS flash lamp.

Detection limit 0.9 ng/mL
Assay range 0.9 to 2000 ng/mL
Incubation time 2h at RT
Specificity IgG1, IgG2, IgG3, IgG4

Miniaturization:

Assay volume can easily be adapted to your assay plate by resizing each volume proportionally, without affecting assay performances. The figure below shows human IgG kit calibration curves in 96 half-well (100µL), 384w small volume (20µL) and 1536w (with 8µL). Readout was on a PHERAstarFS laser.

For more information on HTRF compatible plates and readers, click here: http://www.cisbio.com/drug-discovery/htrf-technology

  96 half-well (100µL) 384w (20µL) 1536w (8µL)
Standard 50 µL 10 µL 4 µL
d2 conjugate 25 µL 5 µL 2 µL
K conjugate 25 µL 5 µL 2 µL

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
Human IgG kit - 500 tests6FHIGPEG
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Human IgG kit - 10,000 tests6FHIGPEH

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