Antibody screening on biochemical targets

A comprehensive line of toolbox reagents for the straightforward development of antibody screening and maturation assays

Several therapeutic antibody action mechanisms have already been described, as illustrated here by the ligand or cytokine blockage approach. This concept applies for the first therapeutic antibody developed for the treatment of inflammatory diseases. Infliximab neutralizes the tumor necrosis factor (TNF) and prevents it from activating its cognate receptor.

Cisbio Bioassays has optimized a comprehensive selection of ready-to-use antibodies and binders coupled to HTRF® fluorescent dyes. This large panel of conjugates enables multiple assays to be developed, and is a flexible approach for addressing a broad variety of therapeutic targets.

Anti-IL6 antibody screening case study

Interleukin 6 (IL6) is a pleiotropic pro-inflammatory cytokine produced by a variety of cell types such as lymphocytes, monocytes, and fibroblasts. The deregulation of IL-6 secretion is involved in several inflammatory processes, e.g. rheumatoid arthritis (RA) and Crohn’s disease. It therefore represents a target of choice in therapeutic antibody development.

Medimmune developed a potent and sensitive antibody by engineering the antibody variable domains using the phage display technology. The description below summarizes the HTRF assay format which they used to screen the ScFv proteins from the phage display selection, based on an immunocompetition.

The assay was built up using a mix of FLAG®-tagged human IL-6 and biotinylated IL-6Rα. The HTRF detection was performed by adding an anti-FLAG IgG labeled with Europium cryptate and Steptavidin XLent! The ScFv proteins were assessed for their capacity to displace the FLAG-hIL6 / biot-IL-6Rα interaction.

Donna K. Finch & al. Whole-Molecule Antibody Engineering: Generation of a High-affinity Anti-IL6 Antibody with Extended Pharmacokinetics. J.Mol. Biol 2011. 411, 791-807.

Anti-IL6 antibody maturation case study

In the second step, Medimmune ran a random and targeted mutagenesis strategy based on Ribosome display and Phage display technologies respectively, in order to increase the affinity and the potency of their antibodies. They selected the biobetter fragments against a reference antibody generated during the screening phase, using a second HTRF format as illustrated here.

The assay was set up using a mix of FLAG®-tagged human IL-6 and biotinylated reference anti-IL-6 IgG1 antibody. The HTRF® detection was performed by adding an anti-FLAG IgG labeled with cryptate and Steptavidin XLent!. The engineered ScFv fragments were assessed for their capacity to displace the FLAG®-hIL6 / Biotin-Anti-IL-6 Reference Ab interaction. Affinity improved ScFv proteins were identified using this biochemical assay format.

Donna K. Finch & al. Whole-Molecule Antibody Engineering: Generation of a High-affinity Anti-IL6 Antibody with Extended Pharmacokinetics. J.Mol. Biol 2011. 411, 791-807.

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
MAb Anti HA-d2 - 5,000 tests610HADAA
MAb Anti HA-d2 - 20,000 tests610HADAB
MAb Anti HA-d2 - 1,000 tests610HADAF
MAb Anti HA-Eu cryptate - 5,000 tests610HAKLA
MAb Anti HA-Eu cryptate - 20,000 tests610HAKLB
MAb Anti HA-Tb cryptate - 5,000 tests610HATAA
MAb Anti HA-Tb cryptate - 20,000 tests610HATAB
MAb Anti HA-XL665 - 5,000 tests610HAXLA
MAb Anti HA-XL665 - 20,000 tests610HAXLB
Streptavidin-d2 - 5,000 tests610SADLA
Streptavidin-d2 - 20,000 tests610SADLB
Streptavidin-Eu cryptate - 5,000 tests610SAKLA
Streptavidin-Eu cryptate - 20,000 tests610SAKLB
Streptavidin-Tb cryptate - 5,000 tests610SATLA
Streptavidin-Tb cryptate - 20,000 tests610SATLB
Streptavidin-XL665 - 5,000 tests610SAXLA
Streptavidin-XL665 - 20,000 tests610SAXLB
MAb Anti FLAG M2-Eu cryptate - 5,000 tests61FG2KLA
MAb Anti FLAG M2-Eu cryptate - 20,000 tests61FG2KLB
MAb Anti FLAG M2-Tb cryptate - 5,000 tests61FG2TLA
MAb Anti FLAG M2-Tb cryptate - 20,000 tests61FG2TLB
MAb Anti FLAG M2-XL665 - 5,000 tests61FG2XLA
MAb Anti FLAG M2-XL665 - 20,000 tests61FG2XLB
MAb Anti GST-d2 - 5,000 tests61GSTDLA
MAb Anti GST-d2 - 20,000 tests61GSTDLB
MAb Anti GST-Eu cryptate - 5,000 tests61GSTKLA
MAb Anti GST-Eu cryptate - 20,000 tests61GSTKLB
MAb Anti GST-Tb cryptate - 5,000 tests61GSTTLA
MAb Anti GST-Tb cryptate - 20,000 tests61GSTTLB
MAb Anti GST-XL665 - 5,000 tests61GSTXLA
MAb Anti GST-XL665 - 20,000 tests61GSTXLB
PAb Anti Human IgG-d2 - 5,000 tests61HFCDAA
PAb Anti Human IgG-Eu cryptate- 5,000 tests61HFCKLA
PAb Anti Human IgG-Eu cryptate - 20,000 tests61HFCKLB
PAb Anti Human IgG-XL665 - 5,000 tests61HFCXLA
PAb Anti Human IgG-XL665 - 20,000 tests61HFCXLB
MAb Anti 6HIS-d2 - 5,000 tests61HISDLA
MAb Anti 6HIS-d2 - 20,000 tests61HISDLB
MAb Anti 6HIS-Eu cryptate - 5,000 tests61HISKLA
MAb Anti 6HIS-Eu cryptate - 20,000 tests61HISKLB
MAb Anti 6HIS-Tb cryptate - 5,000 tests61HISTLA
MAb Anti 6HIS-Tb cryptate - 20,000 tests61HISTLB
MAb Anti 6HIS-XL665 - 5,000 tests61HISXLA
MAb Anti 6HIS-XL665 - 20,000 tests61HISXLB
MAb Anti MBP-d2 - 5,000 tests61MBPDAA
MAb Anti MBP-d2 - 20,000 tests61MBPDAB
MAb Anti MBP-Eu cryptate - 5,000 tests61MBPKAA
MAb Anti MBP-Eu cryptate - 20,000 tests61MBPKAB
MAb Anti MBP-Tb cryptate - 5,000 tests61MBPTAA
MAb Anti MBP-Tb cryptate - 20,000 tests61MBPTAB
MAb Anti cmyc-d2 - 5,000 tests61MYCDAA
MAb Anti cmyc-d2 - 20,000 tests61MYCDAB
MAb Anti cmyc-Eu cryptate - 5,000 tests61MYCKLA
MAb Anti cmyc-Eu cryptate - 20,000 tests61MYCKLB
MAb Anti cmyc-Tb cryptate - 5,000 tests61MYCTAA
MAb Anti cmyc-Tb cryptate - 20,000 tests61MYCTAB
MAb Anti cmyc-XL665 - 5,000 tests61MYCXLA
MAb Anti cmyc-XL665 - 20,000 tests61MYCXLB
PAb Anti Mouse IgG-d2 - 5,000 tests61PAMDAA
PAb Anti Mouse-IgG-d2 - 20,000 tests61PAMDAB
PAb Anti Mouse IgG-Eu cryptate - 5,000 tests61PAMKLA
PAb Anti Mouse IgG-Eu cryptate - 20,000 tests61PAMKLB
PAb Anti Mouse IgG-XL665 - 5,000 tests61PAMXLA
PAb Anti Mouse IgG-XL665 - 20,000 tests61PAMXLB
PAb Anti Rabbit IgG-Eu cryptate - 5,000 tests61PARKLA
PAb Anti Rabbit gG-Eu cryptate - 20,000 tests61PARKLB
PAb Anti Rabbit IgG-XL665 - 5,000 tests61PARXLA
PAb Anti Rabbit IgG-XL666 - 20,000 tests61PARXLB
d2 Labeling kit62D2DPEA
Europium cryptate Labeling kit62EUSPEA

Companion products

DescriptionCat. noProduct insertMSDS
Streptavidin-d2 - 1,000 tests610SADLF
Streptavidin-Tb cryptate - 1,000 tests610SATLF
Streptavidin-XL665 - 1,000 tests610SAXLF

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