Integrated top-down and bottom-up Proteomic Approach to Characterize The Antigen-Binding

An integrated top-down and bottom-up proteomic approach to characterize the antigen-binding fragment of antibodies.

We have previously shown that different individuals exposed to the same antigen produce antibodies with identical mutations in their complementarity determining regions (CDR), suggesting that CDR tryptic peptides can serve as biomarkers for disease diagnosis and prognosis.

Complete Fabs derived from disease-specific antibodies have even higher potential; they could potentially be used for disease treatment and are required to identify the antigens toward which the antibodies are directed. However, complete Fab sequence characterization via LC-MS analysis of tryptic peptides (i.e. bottom-up) has proven to be impractical for mixtures of antibodies.

To tackle this challenge, we have developed an integrated bottom-up and top-down MS approach, employing 2D chromatography coupled with Fourier transform mass spectrometry (FTMS), and applied this approach for full characterization of the variable parts of two pharmaceutical monoclonal antibodies with sensitivity comparable to the bottom-up standard.

These efforts represent an essential step toward the identification of disease specific antibodies in patient samples with potentially significant clinical impact.

Selective 351 nm photodissociation of cysteine-containing peptides for discrimination of antigen-binding regions of IgG fragments in bottom-up liquid chromatography-tandem mass spectrometry workflows.

Despite tremendous inroads in the development of more sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) strategies for mass spectrometry-based proteomics, there remains a significant need for enhancing the selectivity of MS/MS-based workflows for streamlined analysis of complex biological mixtures.

Here, a novel LC-MS/MS platform based on 351 nm ultraviolet photodissociation (UVPD) is presented for the selective analysis of cysteine-peptide subsets in complex protein digests. Cysteine-selective UVPD is mediated through the site-specific conjugation of reduced cysteine residues with a 351 nm active chromogenic Alexa Fluor 350 (AF350) maleimide tag.

Only peptides containing the AF350 chromophore undergo photodissociation into extensive arrays of b- and y-type fragment ions, thus providing a facile means for differentiating cysteine-peptide targets from convoluting peptide backgrounds.

With the use of this approach in addition to strategic proteolysis, the selective analysis of diagnostic heavy-chain complementarity determining regions (CDRs) of single-chain antibody (scAb) fragments is demonstrated.

Fragments of the Roelofs effect: a bottom-up effect equal to the sum of its parts.

The Roelofs effect is a distortion of perceived space that occurs when a large frame whose center is offset left or right from the objective midline is presented visually to an observer and causes a bias in the observer’s subjective judgment of midline.

Experiments were designed to test whether an isolated fragment (left or right end) of a Roelofs-inducing frame was capable of generating the Roelofs effect and to determine whether prior experience with intact frames would provide a top-down influence that would bias the Roelofs effect resulting from fragment presentation.

Although the fragments did induce an effect, top-down information did not play a significant role even after a 5-day training paradigm. Instead, we found that the effect generated by an intact frame was equal in magnitude to the sum of the effects generated by the individual fragments.

In addition, perception was found to be differentially affected by the two ends of the frame, with fragments falling in the right visual field causing a larger effect than those falling in the left.

Monkey Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E09R0362-96	BlueGene	1 plate of 96 wells	EUR 685
Description: A competitive ELISA for quantitative measurement of Monkey Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Dog Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E08R0362-192T	BlueGene	192 tests	EUR 1270
Description: A competitive ELISA for quantitative measurement of Canine Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Dog Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E08R0362-48	BlueGene	1 plate of 48 wells	EUR 520
Description: A competitive ELISA for quantitative measurement of Canine Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Dog Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E08R0362-96	BlueGene	1 plate of 96 wells	EUR 685
Description: A competitive ELISA for quantitative measurement of Canine Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Human Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E01R0362-192T	BlueGene	192 tests	EUR 1270
Description: A competitive ELISA for quantitative measurement of Human Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Human Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E01R0362-48	BlueGene	1 plate of 48 wells	EUR 520
Description: A competitive ELISA for quantitative measurement of Human Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Human Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E01R0362-96	BlueGene	1 plate of 96 wells	EUR 685
Description: A competitive ELISA for quantitative measurement of Human Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Pig Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E07R0362-192T	BlueGene	192 tests	EUR 1270
Description: A competitive ELISA for quantitative measurement of Porcine Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Pig Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E07R0362-48	BlueGene	1 plate of 48 wells	EUR 520
Description: A competitive ELISA for quantitative measurement of Porcine Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Pig Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E07R0362-96	BlueGene	1 plate of 96 wells	EUR 685
Description: A competitive ELISA for quantitative measurement of Porcine Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Rabbit Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E04R0362-192T	BlueGene	192 tests	EUR 1270
Description: A competitive ELISA for quantitative measurement of Rabbit Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Rabbit Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E04R0362-48	BlueGene	1 plate of 48 wells	EUR 520
Description: A competitive ELISA for quantitative measurement of Rabbit Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Rabbit Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E04R0362-96	BlueGene	1 plate of 96 wells	EUR 685
Description: A competitive ELISA for quantitative measurement of Rabbit Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Goat Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E06R0362-192T	BlueGene	192 tests	EUR 1270
Description: A competitive ELISA for quantitative measurement of Goat Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Goat Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E06R0362-48	BlueGene	1 plate of 48 wells	EUR 520
Description: A competitive ELISA for quantitative measurement of Goat Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Goat Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E06R0362-96	BlueGene	1 plate of 96 wells	EUR 685
Description: A competitive ELISA for quantitative measurement of Goat Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Mouse Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E03R0362-192T	BlueGene	192 tests	EUR 1270
Description: A competitive ELISA for quantitative measurement of Mouse Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Mouse Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E03R0362-48	BlueGene	1 plate of 48 wells	EUR 520
Description: A competitive ELISA for quantitative measurement of Mouse Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Mouse Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E03R0362-96	BlueGene	1 plate of 96 wells	EUR 685
Description: A competitive ELISA for quantitative measurement of Mouse Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Guinea pig Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E05R0362-192T	BlueGene	192 tests	EUR 1270
Description: A competitive ELISA for quantitative measurement of Guinea pig Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Guinea pig Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E05R0362-48	BlueGene	1 plate of 48 wells	EUR 520
Description: A competitive ELISA for quantitative measurement of Guinea pig Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Guinea pig Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
E05R0362-96	BlueGene	1 plate of 96 wells	EUR 685
Description: A competitive ELISA for quantitative measurement of Guinea pig Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Recombinant Aspergillus Oryzae ACT1 Protein (aa 1-375) [His]
VAng-Wyb3947-1mg	Creative Biolabs	1 mg	EUR 4702
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Actin, recombinant protein.

Recombinant Aspergillus Oryzae ADH5 Protein (aa 1-383) [His]
VAng-Wyb3948-1mg	Creative Biolabs	1 mg	EUR 4730
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Alcohol Dehydrogenase 5 (Class III), chi Polypeptide, recombinant protein.

Recombinant Aspergillus Oryzae Amy3 Protein (aa 22-499) [His]
VAng-Wyb3949-1mg	Creative Biolabs	1 mg	EUR 5109
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) alpha-Amylase A Type-3, recombinant protein.

Recombinant Aspergillus Oryzae AO090103000074 Protein (aa 19-316) [His]
VAng-Wyb3950-1mg	Creative Biolabs	1 mg	EUR 4397
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Tannase, recombinant protein.

Recombinant Aspergillus Oryzae ATG12 Protein (aa 1-176) [His]
VAng-Wyb3951-1mg	Creative Biolabs	1 mg	EUR 3916
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Autophagy Related 12, recombinant protein.

Recombinant Aspergillus Oryzae ATG3 Protein (aa 1-356) [His]
VAng-Wyb3952-1mg	Creative Biolabs	1 mg	EUR 4625
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Autophagy Related 3 Homolog, recombinant protein.

Recombinant Aspergillus Oryzae ATG5 Protein (aa 1-322) [His]
VAng-Wyb3953-1mg	Creative Biolabs	1 mg	EUR 4494
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Autophagy Related 5 Homolog, recombinant protein.

Recombinant Aspergillus Oryzae BCP1 Protein (aa 1-290) [His]
VAng-Wyb3954-1mg	Creative Biolabs	1 mg	EUR 4364
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Bcp1p, recombinant protein.

Recombinant Aspergillus Oryzae BUD32 Protein (aa 1-272) [His]
VAng-Wyb3955-1mg	Creative Biolabs	1 mg	EUR 4292
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Bud32p, recombinant protein.

Recombinant Aspergillus Oryzae CAP1 Protein (aa 1-273) [His]
VAng-Wyb3956-1mg	Creative Biolabs	1 mg	EUR 4298
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Cap1p, recombinant protein.

Recombinant Aspergillus Oryzae CBP4 Protein (aa 1-116) [His]
VAng-Wyb3958-1mg	Creative Biolabs	1 mg	EUR 3677
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Cbp4p, recombinant protein.

Recombinant Aspergillus Oryzae CFD1 Protein (aa 1-315) [His]
VAng-Wyb3959-1mg	Creative Biolabs	1 mg	EUR 4466
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Cfd1p, recombinant protein.

Recombinant Aspergillus Oryzae CHZ1 Protein (aa 1-110) [His]
VAng-Wyb3960-1mg	Creative Biolabs	1 mg	EUR 3655
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Chz1p, recombinant protein.

Recombinant Aspergillus Oryzae CTU2 Protein (aa 1-361) [His]
VAng-Wyb3962-1mg	Creative Biolabs	1 mg	EUR 4645
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Cytosolic Thiouridylase Subunit 2 Homolog, recombinant protein.

Recombinant Aspergillus Oryzae DDI1 Protein (aa 1-402) [His]
VAng-Wyb3963-1mg	Creative Biolabs	1 mg	EUR 4807
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) DNA-Damage Inducible 1 Homolog 1, recombinant protein.

Recombinant Aspergillus Oryzae DML1 Protein (aa 1-493) [His]
VAng-Wyb3964-1mg	Creative Biolabs	1 mg	EUR 5166
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Dml1p, recombinant protein.

Recombinant Aspergillus Oryzae DOT1L Protein (aa 1-335) [His]
VAng-Wyb3965-1mg	Creative Biolabs	1 mg	EUR 4540
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) DOT1-Like, Histone H3 Methyltransferase, recombinant protein.

Recombinant Aspergillus Oryzae DRE2 Protein (aa 1-313) [His]
VAng-Wyb3966-1mg	Creative Biolabs	1 mg	EUR 4455
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Dre2p, recombinant protein.

Recombinant Aspergillus Oryzae EFG1 Protein (aa 1-320) [His]
VAng-Wyb3967-1mg	Creative Biolabs	1 mg	EUR 4482
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Efg1p, recombinant protein.

Recombinant Aspergillus Oryzae Egd2 Protein (aa 1-202) [His]
VAng-Wyb3968-1mg	Creative Biolabs	1 mg	EUR 4017
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Nascent Polypeptide-Associated Complex Subunit alpha, recombinant protein.

Recombinant Aspergillus Oryzae EIF3E Protein (aa 1-451) [His]
VAng-Wyb3969-1mg	Creative Biolabs	1 mg	EUR 4999
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Eukaryotic Translation Initiation Factor 3 Subunit E, recombinant protein.

Recombinant Aspergillus Oryzae EIF3G Protein (aa 1-287) [His]
VAng-Wyb3970-1mg	Creative Biolabs	1 mg	EUR 4350
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Eukaryotic Translation Initiation Factor 3, Subunit G, recombinant protein.

Recombinant Aspergillus Oryzae EIF3I Protein (aa 1-335) [His]
VAng-Wyb3971-1mg	Creative Biolabs	1 mg	EUR 4540
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Eukaryotic Translation Initiation Factor 3, Subunit I, recombinant protein.

Recombinant Aspergillus Oryzae ESF2 Protein (aa 1-335) [His]
VAng-Wyb3972-1mg	Creative Biolabs	1 mg	EUR 4540
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Esf2p, recombinant protein.

Recombinant Aspergillus Oryzae FIS1 Protein (aa 1-126) [His]
VAng-Wyb3973-1mg	Creative Biolabs	1 mg	EUR 3716
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Fis1p, recombinant protein.

Recombinant Aspergillus Oryzae FPR1 Protein (aa 1-116) [His]
VAng-Wyb3974-1mg	Creative Biolabs	1 mg	EUR 3677
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Fpr1p, recombinant protein.

Recombinant Aspergillus Oryzae FPR2 Protein (aa 20-134) [His]
VAng-Wyb3975-1mg	Creative Biolabs	1 mg	EUR 3673
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Fpr2p, recombinant protein.

Recombinant Aspergillus Oryzae FPR4 Protein (aa 1-470) [His]
VAng-Wyb3976-1mg	Creative Biolabs	1 mg	EUR 5076
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Fpr4p, recombinant protein.

Recombinant Aspergillus Oryzae GAPDH Protein (aa 1-338) [His]
VAng-Wyb3977-1mg	Creative Biolabs	1 mg	EUR 4553
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Glyceraldehyde-3-Phosphate Dehydrogenase, recombinant protein.

Recombinant Aspergillus Oryzae GCD14 Protein (aa 1-474) [His]
VAng-Wyb3978-1mg	Creative Biolabs	1 mg	EUR 5090
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Gcd14p, recombinant protein.

Recombinant Aspergillus Oryzae GINS2 Protein (aa 1-269) [His]
VAng-Wyb3979-1mg	Creative Biolabs	1 mg	EUR 4278
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) GINS Complex Subunit 2 (Psf2 Homolog), recombinant protein.

Recombinant Aspergillus Oryzae H2A Protein (aa 2-133) [His]
VAng-Wyb3980-1mg	Creative Biolabs	1 mg	EUR 3740
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Histone H2A, recombinant protein.

Recombinant Aspergillus Oryzae HCR1 Protein (aa 1-265) [His]
VAng-Wyb3981-1mg	Creative Biolabs	1 mg	EUR 4266
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) COenzyme Q (Ubiquinone) Hcr1p, recombinant protein.

Recombinant Aspergillus Oryzae JIP5 Protein (aa 1-417) [His]
VAng-Wyb3982-1mg	Creative Biolabs	1 mg	EUR 4867
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Jip5p, recombinant protein.

Recombinant Aspergillus Oryzae KAE1 Protein (aa 1-358) [His]
VAng-Wyb3983-1mg	Creative Biolabs	1 mg	EUR 4636
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Kae1p, recombinant protein.

Recombinant Aspergillus Oryzae LCL2 Protein (aa 22-1226) [His]
VAng-Wyb3984-1mg	Creative Biolabs	1 mg	EUR 3616
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Lcl2p, recombinant protein.

Recombinant Aspergillus Oryzae LIA1 Protein (aa 1-339) [His]
VAng-Wyb3985-1mg	Creative Biolabs	1 mg	EUR 4559
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Lia1p, recombinant protein.

Recombinant Aspergillus Oryzae LOC1 Protein (aa 1-190) [His]
VAng-Wyb3986-1mg	Creative Biolabs	1 mg	EUR 3971
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) COenzyme Q (Ubiquinone) Loc1p, recombinant protein.

Recombinant Aspergillus Oryzae LOC100799412 Protein (aa 1-344) [His]
VAng-Wyb3987-1mg	Creative Biolabs	1 mg	EUR 4578
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Mitogen-Activated Protein Kinase MpkC, recombinant protein.

Recombinant Aspergillus Oryzae MDE1 Protein (aa 1-241) [His]
VAng-Wyb3988-1mg	Creative Biolabs	1 mg	EUR 4172
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Mde1p, recombinant protein.

Recombinant Aspergillus Oryzae MED21 Protein (aa 1-208) [His]
VAng-Wyb3989-1mg	Creative Biolabs	1 mg	EUR 4042
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Mediator Complex Subunit 21, recombinant protein.

Recombinant Aspergillus Oryzae MED6 Protein (aa 1-328) [His]
VAng-Wyb3990-1mg	Creative Biolabs	1 mg	EUR 4518
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Mediator Complex Subunit 6, recombinant protein.

Recombinant Aspergillus Oryzae MED7 Protein (aa 1-261) [His]
VAng-Wyb3991-1mg	Creative Biolabs	1 mg	EUR 4251
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Mediator Complex Subunit 7, recombinant protein.

Recombinant Aspergillus Oryzae MED8 Protein (aa 1-244) [His]
VAng-Wyb3992-1mg	Creative Biolabs	1 mg	EUR 4185
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Mediator Complex Subunit 8, recombinant protein.

Recombinant Aspergillus Oryzae METTL1 Protein (aa 1-330) [His]
VAng-Wyb3993-1mg	Creative Biolabs	1 mg	EUR 4521
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Methyltransferase Like 1, recombinant protein.

Recombinant Aspergillus Oryzae MMM1 Protein (aa 47-494) [His]
VAng-Wyb3994-1mg	Creative Biolabs	1 mg	EUR 4991
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Mmm1p, recombinant protein.

Recombinant Aspergillus Oryzae MOCS2 Protein (aa 1-179) [His]
VAng-Wyb3995-1mg	Creative Biolabs	1 mg	EUR 3924
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Molybdenum Cofactor Synthesis 2, recombinant protein.

Recombinant Aspergillus Oryzae Mri1 Protein (aa 1-381) [His]
VAng-Wyb3996-1mg	Creative Biolabs	1 mg	EUR 4724
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Methylthioribose-1-Phosphate Isomerase Homolog, recombinant protein.

Recombinant Aspergillus Oryzae MZM1 Protein (aa 19-114) [His]
VAng-Wyb3997-1mg	Creative Biolabs	1 mg	EUR 3597
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Mzm1p, recombinant protein.

Recombinant Aspergillus Oryzae NBP35 Protein (aa 1-325) [His]
VAng-Wyb3998-1mg	Creative Biolabs	1 mg	EUR 4502
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Nbp35p, recombinant protein.

Recombinant Aspergillus Oryzae NGLY1 Protein (aa 1-457) [His]
VAng-Wyb3999-1mg	Creative Biolabs	1 mg	EUR 5024
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) N-Glycanase 1, recombinant protein.

Recombinant Aspergillus Oryzae NUCS Protein (aa 21-287) [His]
VAng-Wyb4000-1mg	Creative Biolabs	1 mg	EUR 4276
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) NucS Gene Product, recombinant protein.

Recombinant Aspergillus Oryzae NUF2 Protein (aa 1-463) [His]
VAng-Wyb4001-1mg	Creative Biolabs	1 mg	EUR 5048
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Spindle Pole Body Protein, recombinant protein.

Recombinant Aspergillus Oryzae P4HB Protein (aa 21-515) [His]
VAng-Wyb4002-1mg	Creative Biolabs	1 mg	EUR 5175
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Prolyl 4-Hydroxylase, beta Polypeptide, recombinant protein.

Recombinant Aspergillus Oryzae PNO1 Protein (aa 1-260) [His]
VAng-Wyb4003-1mg	Creative Biolabs	1 mg	EUR 4246
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Pno1p, recombinant protein.

Recombinant Aspergillus Oryzae POA1 Protein (aa 1-201) [His]
VAng-Wyb4004-1mg	Creative Biolabs	1 mg	EUR 4015
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Poa1p, recombinant protein.

Recombinant Aspergillus Oryzae PPME1 Protein (aa 1-427) [His]
VAng-Wyb4005-1mg	Creative Biolabs	1 mg	EUR 4906
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Protein Phosphatase Methylesterase 1, recombinant protein.

Recombinant Aspergillus Oryzae PXR1 Protein (aa 1-306) [His]
VAng-Wyb4006-1mg	Creative Biolabs	1 mg	EUR 4427
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Pxr1p, recombinant protein.

Recombinant Aspergillus Oryzae RBBP7 Protein (aa 1-436) [His]
VAng-Wyb4007-1mg	Creative Biolabs	1 mg	EUR 4944
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Retinoblastoma Binding Protein 7, recombinant protein.

Recombinant Aspergillus Oryzae RMT2 Protein (aa 1-413) [His]
VAng-Wyb4008-1mg	Creative Biolabs	1 mg	EUR 4848
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Rmt2p, recombinant protein.

Recombinant Aspergillus Oryzae RPS1 Protein (aa 2-256) [His]
VAng-Wyb4009-1mg	Creative Biolabs	1 mg	EUR 4227
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Rps1 Gene Product, recombinant protein.

Recombinant Aspergillus Oryzae RRD1 Protein (aa 1-478) [His]
VAng-Wyb4010-1mg	Creative Biolabs	1 mg	EUR 5109
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Rrd1p, recombinant protein.

Recombinant Aspergillus Oryzae RRD2 Protein (aa 1-422) [His]
VAng-Wyb4011-1mg	Creative Biolabs	1 mg	EUR 4887
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Rrd2p, recombinant protein.

Recombinant Aspergillus Oryzae RRG9 Protein (aa 54-284) [His]
VAng-Wyb4012-1mg	Creative Biolabs	1 mg	EUR 4133
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Rrg9p, recombinant protein.

Recombinant Aspergillus Oryzae RRP36 Protein (aa 1-352) [His]
VAng-Wyb4013-1mg	Creative Biolabs	1 mg	EUR 4612
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Rrp36p, recombinant protein.

Recombinant Aspergillus Oryzae SFH5 Protein (aa 1-455) [His]
VAng-Wyb4014-1mg	Creative Biolabs	1 mg	EUR 5015
Description: Aspergillus Oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold) Sfh5p, recombinant protein.

Induction of cell cycle arrest and apoptosis by tomentosin in hepatocellular carcinoma HepG2 and Huh7 cells.

Tomentosin, a sesquiterpene lactone, is known to possess various biological activities. However, its anticarcinogenic activity against human hepatocellular carcinoma (HCC) cells has not been investigated in detail. Thus, this study aimed to elucidate the cytotoxic mechanism of tomentosin in human HCC cell lines HepG2 and Huh7.

WST-1, cell counting, and colony formation assay results showed that treatment with tomentosin decreased the viability and suppressed the proliferation rate of HepG2 and Huh7 cells in a dose- and time-dependent manner. Cell cycle analysis revealed increased population of cells at the SubG1 and G2/M stage, and decreased population of cells at the G0/1 stage in HepG2 and Huh7 cells treated with tomentosin.

Annexin V/propidium iodide double staining and TUNEL assay results showed increased apoptotic cell population and DNA fragmentation in HepG2 and Huh7 cells treated with tomentosin.

Western blotting analysis results showed that tomentosin treatment significantly increased the expression level of Bax, Bim (short form), cleaved PARP1, FOXO3, p53, pSer15p53, pSer20p53, pSer46p53, p21, and p27, but decreased the expression of Bcl2, caspase3, caspase7, caspase9, cyclin-dependent kinase 2 (CDK2), CDK4, CDK6, cyclinB1, cyclinD1, cyclinD2, cyclinD3, and cyclinE in a dose-dependent manner.

Taken together, this study revealed that tomentosin, which acted through cell cycle arrest and apoptosis, may be a useful therapeutic option against HCC.

Keywords: FOXO3; Tomentosin; apoptosis; cell cycle arrest; hepatocellular carcinoma; p53.