Reaction #1121739

ord-34732e463156470ea025a3732cc53098

Reaction equation

O=C(O)CCCC[C@@H]1SC[C@@H]2NC(=O)N[C@@H]21.O=C1C=CC(=O)N1
biotin-maleimide
N[C@@H](CS)C(=O)O
cysteine
N[C@@H](CS)C(=O)O
cysteine
→
O=C(O)CCCC[C@@H]1SC[C@@H]2NC(=O)N[C@@H]21
biotin
Yield 100.0%

Reactants

—
thiol
O=C(O)CCCC[C@@H]1SC[C@@H]2NC(=O)N[C@@H]21.O=C1C=CC(=O)N1
biotin-maleimide
N[C@@H](CS)C(=O)O
cysteine
—
thiol
—
cysteine amino acid
—
thiol
—
cysteine amino acid
—
thiol
—
cysteine amino acids
N[C@@H](CS)C(=O)O
cysteine

Reagents

None

Conditions

Detailed conditions
See reaction.notes.procedure_details.

Procedure

The term “thiol reactivity value” is a quantitative characterization of the reactivity of free cysteine amino acids. The thiol reactivity value is the percentage of a free cysteine amino acid in a cysteine engineered antibody which reacts with a thiol-reactive reagent, and converted to a maximum value of 1. For example, a free cysteine amino acid on a cysteine engineered antibody which reacts in 100% yield with a thiol-reactive reagent, such as a biotin-maleimide reagent, to form a biotin-labelled antibody has a thiol reactivity value of 1.0. Another cysteine amino acid engineered into the same or different parent antibody which reacts in 80% yield with a thiol-reactive reagent has a thiol reactivity value of 0.8. Another cysteine amino acid engineered into the same or different parent antibody which fails totally to react with a thiol-reactive reagent has a thiol reactivity value of 0. Determination of the thiol reactivity value of a particular cysteine may be conducted by ELISA assay, mass spectroscopy, liquid chromatography, autoradiography, or other quantitative analytical tests.

Source

DOI: 10.6084/m9.figshare.5104873.v1Patent: US08545850B2uspto-grants-2013_10