Literature summary for 1.13.12.6 extracted from

Wu, C.; Mino, K.; Akimoto, H.; Kawabata, M.; Nakamura, K.; Ozaki, M.; Ohmiya, Y.
In vivo far-red luminescence imaging of a biomarker based on BRET from Cypridina bioluminescence to an organic dye (2009), Proc. Natl. Acad. Sci. USA, 106, 15599-15603.

Search for more literature for 1.13.12.6

Application

Application	Comment	Organism
diagnostics	far-red luminescence imaging technology to visualize tumor specific antigens on cell surfaces in vitro (human hepato-cellular carcinoma cell line Huh-7) and in living bodies (mice xenografted with human Delta-like protein-positive Huh-7 cells), based on a far-red fluorescent indocyanine derivative conjugated to biotinylated Cypridina luciferase linked to an anti-human Delta-like protein (Dlk-1) monoclonal antibody (embryonic antigens expressed on the surface of many cancer cells) via biotin-avidin interaction	Cypridina noctiluca

Cloned(Commentary)

Cloned (Comment)	Organism
expression of human Delta-like protein (Dlk-1) antigen in HEK-293 cells, immunization of mice (BALB/c) with HEK293-Dlk-1 cells or with the expression vector that encodes full-length Dlk-1 cDNA	Cypridina noctiluca

Protein Variants

Protein Variants	Comment	Organism
additional information	biotynilation of the enzyme by attachment of an Avi-tag of a 16-residue peptide to the C-terminus of the luciferase, conjugation to the indocyanine derivative HiLyte Fluor 647 hydrazine via the glycol-chains of the enzyme	Cypridina noctiluca

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
200000	-	SDS-PAGE, reduced antibody-avidin conjugate of the conjugated enzyme (FBP-IgG)	Cypridina noctiluca

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
Cypridina luciferin + O2	Cypridina noctiluca	-	oxidized Cypridina luciferin + CO2 + hv	conjugated enzyme exhibits bimodal bioluminescence spectrum at 460 nm and 675 nm, exhibits higher light intensity in mouse blood than unconjugated enzyme, far-red emission peak dominates in blood while the 460 nm peak dominates in buffers	?

Organism

Organism	UniProt	Comment	Textmining
Cypridina noctiluca	-	sea-firefly	-

Purification (Commentary)

Purification (Comment)	Organism
biotinylated enzyme is incubated with 1 microM sodium metaperiodate in 0.1 M sodium acetate buffer (pH 5.2), reaction stopped by glycerol addition, loading onto a size exclusion column, oxidized biotinylated enzyme eluted with the same buffer, pooling of bioluminescent fractions, concentration by ultrafree-0.5 centrifugal filter device, addition of 10 mM HiLyte Fluor 647 hydrazine in 0.1 M sodium acetate buffer (pH 5.2), after incubation loading onto a size-exclusion column, elution with 0.1 M potassium phosphate buffer (pH 7.2) containing 0.15 M NaCl. Treatment of human Delta-like protein (Dlk-1) antibody with 2-mercaptoethylamine (cleaving hinge-region disulfide bonds between heavy chains of antibody molecules) and conjugation of half antibodies to maleimide-activated avidin and purification on a size-exclusion column results in a 200 kDa conjugate	Cypridina noctiluca

Purification (Comment)

Organism

biotinylated enzyme is incubated with 1 microM sodium metaperiodate in 0.1 M sodium acetate buffer (pH 5.2), reaction stopped by glycerol addition, loading onto a size exclusion column, oxidized biotinylated enzyme eluted with the same buffer, pooling of bioluminescent fractions, concentration by ultrafree-0.5 centrifugal filter device, addition of 10 mM HiLyte Fluor 647 hydrazine in 0.1 M sodium acetate buffer (pH 5.2), after incubation loading onto a size-exclusion column, elution with 0.1 M potassium phosphate buffer (pH 7.2) containing 0.15 M NaCl. Treatment of human Delta-like protein (Dlk-1) antibody with 2-mercaptoethylamine (cleaving hinge-region disulfide bonds between heavy chains of antibody molecules) and conjugation of half antibodies to maleimide-activated avidin and purification on a size-exclusion column results in a 200 kDa conjugate

Cypridina noctiluca

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
Cypridina luciferin + O2	-	Cypridina noctiluca	oxidized Cypridina luciferin + CO2 + hv	conjugated enzyme exhibits bimodal bioluminescence spectrum at 460 nm and 675 nm, exhibits higher light intensity in mouse blood than unconjugated enzyme, far-red emission peak dominates in blood while the 460 nm peak dominates in buffers	?

Synonyms

Synonyms	Comment	Organism
CLuc	-	Cypridina noctiluca
Cypridina luciferase	-	Cypridina noctiluca
Fbp	biotinylated Cypridina luciferase conjugated to far-red fluorescent indocyanine derivative	Cypridina noctiluca
FBP-IgG	conjugate of reduced monoclonal antibody to conjugated enzyme FBP	Cypridina noctiluca

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.5	8	no difference in normalized bioluminescence spectra of conjugated enzyme in 100 mM sodium phosphate buffer (pH 6.5), 100 mM Tris-HCl buffer (pH 7.4) or 100 mM Tris-HCl buffer (pH 8) each containing 0.1 M NaCl	Cypridina noctiluca