Capillary isoelectric focusing (cIEF) was online coupled to a Q-TOF MS by a flow-through microvial interface for the analysis of therapeutic mAb.
Intact molecular weights obtained from the mass spectrum deconvolution of separated charge variants provided information on the structural heterogeneity of therapeutic mAbs. A sandwich cIEF-MS configuration composed of anolyte, sample, and catholyte segments sequentially injected into a neutrally coated capillary was used for the charge heterogeneity separation of four mAbs.
Acetic acid and ammonium hydroxide were used in places of the non-volatile acids and bases commonly used for IEF but are incompatible with online MS detection. Glycerol was added as the anti-convective reagent.
5-BrdU |
B3589-5.1 |
ApexBio |
10 mM (in 1mL DMSO) |
EUR 113 |
Description: 5-Bromodeoxyuridine (5-BrdU) is an analog of Thy that is incorporated into the DNA of cells undergoing the S-phase and can be detected by monoclonal antibodies or polyclona111. |
5-BrdU |
B3589-500 |
ApexBio |
500 mg |
EUR 108 |
Description: 5-Bromodeoxyuridine (5-BrdU) is an analog of Thy that is incorporated into the DNA of cells undergoing the S-phase and can be detected by monoclonal antibodies or polyclona111. |
BRDU antibody |
20R-BS001 |
Fitzgerald |
250 ug |
EUR 553 |
Description: Sheep polyclonal BRDU antibody |
BRDU antibody |
10R-6763 |
Fitzgerald |
1 ml |
EUR 402 |
Description: Mouse monoclonal BRDU antibody |
BRDU antibody |
10R-7945 |
Fitzgerald |
100 ug |
EUR 370 |
Description: Mouse monoclonal BRDU antibody |
BRDU antibody |
10R-B121a |
Fitzgerald |
100 ug |
EUR 445 |
Description: Mouse monoclonal BRDU antibody |
BRDU antibody |
20-BS17 |
Fitzgerald |
250 ug |
EUR 386 |
Description: Sheep polyclonal BRDU antibody |
BRDU antibody |
10R-2321 |
Fitzgerald |
1 mL |
EUR 601 |
Description: Mouse monoclonal BRDU antibody |
BRDU antibody |
10R-2356 |
Fitzgerald |
1.5 mL |
EUR 490 |
Description: Mouse monoclonal BRDU antibody |
Sheep IS ELISA Kit |
ESI0046 |
Abclonal |
96Tests |
EUR 521 |
Anserini IS ELISA Kit |
EAI0046 |
Abclonal |
96Tests |
EUR 521 |
Rat IS ELISA Kit |
ERI0046 |
Abclonal |
96Tests |
EUR 521 |
Rabbit IS ELISA Kit |
ERTI0046 |
Abclonal |
96Tests |
EUR 521 |
Mouse IS ELISA Kit |
EMI0046 |
Abclonal |
96Tests |
EUR 521 |
Monkey IS ELISA Kit |
EMKI0046 |
Abclonal |
96Tests |
EUR 521 |
Porcine IS ELISA Kit |
EPI0046 |
Abclonal |
96Tests |
EUR 521 |
Human IS ELISA Kit |
EHI0046 |
Abclonal |
96Tests |
EUR 521 |
Canine IS ELISA Kit |
ECI0046 |
Abclonal |
96Tests |
EUR 521 |
Chicken IS ELISA Kit |
ECKI0046 |
Abclonal |
96Tests |
EUR 521 |
Bovine IS ELISA Kit |
EBI0046 |
Abclonal |
96Tests |
EUR 521 |
Goat IS ELISA Kit |
EGTI0046 |
Abclonal |
96Tests |
EUR 521 |
Anti-BrdU Purified |
11-286-C025 |
ExBio |
0.025 mg |
EUR 108 |
Anti-BrdU Purified |
11-286-C100 |
ExBio |
0.1 mg |
EUR 177 |
Anti-BrdU Purified |
11-682-C025 |
ExBio |
0.025 mg |
EUR 108 |
Anti-BrdU Purified |
11-682-C100 |
ExBio |
0.1 mg |
EUR 177 |
Anti-BrdU PE |
1P-682-T025 |
ExBio |
25 tests |
EUR 140 |
Anti-BrdU PE |
1P-682-T100 |
ExBio |
100 tests |
EUR 240 |
Bromodeoxyuridine (BrdU) Antibody |
abx000064-100ul |
Abbexa |
100 ul |
EUR 411 |
|
BrdU Mouse mAb |
A1482-100ul |
Abclonal |
100 ul |
EUR 308 |
BrdU Mouse mAb |
A1482-200ul |
Abclonal |
200 ul |
Ask for price |
BrdU Mouse mAb |
A1482-20ul |
Abclonal |
20 ul |
Ask for price |
BrdU Mouse mAb |
A1482-50ul |
Abclonal |
50 ul |
EUR 223 |
Bromodeoxyuridine (BrdU) Antibody |
abx412356-50ug |
Abbexa |
50 ug |
EUR 509 |
|
Bromodeoxyuridine (BrdU) Antibody |
abx414257-02mg |
Abbexa |
0.2 mg |
EUR 565 |
|
Bromodeoxyuridine (BrdU) Antibody |
abx414259-01mg |
Abbexa |
0.1 mg |
EUR 439 |
|
Bromodeoxyuridine (BrdU) Antibody |
abx414260-20ug |
Abbexa |
20 ug |
EUR 272 |
|
Bromodeoxyuridine (BrdU) Antibody |
abx230952-100ug |
Abbexa |
100 ug |
EUR 509 |
|
anti- BrdU antibody |
FNab00952 |
FN Test |
100µg |
EUR 548.75 |
|
Description: Antibody raised against BrdU |
BrdU (Bu20a) Antibody |
48275-100ul |
SAB |
100ul |
EUR 333 |
BrdU (Bu20a) Antibody |
48275-50ul |
SAB |
50ul |
EUR 239 |
Anti-BrdU Antibody |
A1284-100 |
Biovision |
|
EUR 398 |
BRDU antibody (FITC) |
61R-1900 |
Fitzgerald |
1 ml |
EUR 409 |
Description: Mouse monoclonal BRDU antibody (FITC) |
Guinea Pig IS ELISA Kit |
EGI0046 |
Abclonal |
96Tests |
EUR 521 |
Sheep BrDU ELISA Kit |
ESB0693 |
Abclonal |
96Tests |
EUR 521 |
Mouse BrDU ELISA Kit |
EMB0693 |
Abclonal |
96Tests |
EUR 521 |
Anserini BrDU ELISA Kit |
EAB0693 |
Abclonal |
96Tests |
EUR 521 |
Rabbit BrDU ELISA Kit |
ERTB0693 |
Abclonal |
96Tests |
EUR 521 |
Monkey BrDU ELISA Kit |
EMKB0693 |
Abclonal |
96Tests |
EUR 521 |
Porcine BrDU ELISA Kit |
EPB0693 |
Abclonal |
96Tests |
EUR 521 |
Rat BrDU ELISA Kit |
ERB0693 |
Abclonal |
96Tests |
EUR 521 |
Human BrDU ELISA Kit |
EHB0693 |
Abclonal |
96Tests |
EUR 521 |
Canine BrDU ELISA Kit |
ECB0693 |
Abclonal |
96Tests |
EUR 521 |
Chicken BrDU ELISA Kit |
ECKB0693 |
Abclonal |
96Tests |
EUR 521 |
Goat BrDU ELISA Kit |
EGTB0693 |
Abclonal |
96Tests |
EUR 521 |
Bovine BrDU ELISA Kit |
EBB0693 |
Abclonal |
96Tests |
EUR 521 |
BrdU (Bromodeoxyuridine)(BRD469) Antibody |
BNUM0469-50 |
Biotium |
50uL |
EUR 395 |
Description: Primary antibody against BrdU (Bromodeoxyuridine)(BRD469), 1mg/mL |
BrdU (Bromodeoxyuridine)(BRD494) Antibody |
BNUM0494-50 |
Biotium |
50uL |
EUR 395 |
Description: Primary antibody against BrdU (Bromodeoxyuridine)(BRD494), 1mg/mL |
BrdU (Bromodeoxyuridine)(BU20a) Antibody |
BNUM0522-50 |
Biotium |
50uL |
EUR 395 |
Description: Primary antibody against BrdU (Bromodeoxyuridine)(BU20a), 1mg/mL |
BrdU (Bromodeoxyuridine)(BRD469) Antibody |
BNUB0469-100 |
Biotium |
100uL |
EUR 209 |
Description: Primary antibody against BrdU (Bromodeoxyuridine)(BRD469), Concentration: 0.2mg/mL |
BrdU (Bromodeoxyuridine)(BRD469) Antibody |
BNUB0469-500 |
Biotium |
500uL |
EUR 458 |
Description: Primary antibody against BrdU (Bromodeoxyuridine)(BRD469), Concentration: 0.2mg/mL |
BrdU (Bromodeoxyuridine)(BRD494) Antibody |
BNUB0494-100 |
Biotium |
100uL |
EUR 209 |
Description: Primary antibody against BrdU (Bromodeoxyuridine)(BRD494), Concentration: 0.2mg/mL |
BrdU (Bromodeoxyuridine)(BRD494) Antibody |
BNUB0494-500 |
Biotium |
500uL |
EUR 458 |
Description: Primary antibody against BrdU (Bromodeoxyuridine)(BRD494), Concentration: 0.2mg/mL |
BrdU (Bromodeoxyuridine)(BU20a) Antibody |
BNUB0522-100 |
Biotium |
100uL |
EUR 209 |
Description: Primary antibody against BrdU (Bromodeoxyuridine)(BU20a), Concentration: 0.2mg/mL |
BrdU (Bromodeoxyuridine)(BU20a) Antibody |
BNUB0522-500 |
Biotium |
500uL |
EUR 458 |
Description: Primary antibody against BrdU (Bromodeoxyuridine)(BU20a), Concentration: 0.2mg/mL |
Bromodeoxyuridine (BrdU) Antibody (FITC) |
abx414258-50ug |
Abbexa |
50 ug |
EUR 342 |
|
Bromodeoxyuridine (BrdU) Antibody (FITC) |
abx201100-100tests |
Abbexa |
100 tests |
EUR 620 |
|
BrdU (Bu20a) Conjugated Antibody |
C48275 |
SAB |
100ul |
EUR 397 |
Human Serum Indoxyl Sulfate (IS) ELISA Kit |
abx257961-96tests |
Abbexa |
96 tests |
EUR 746 |
|
Human IS(serum indoxyl sulfate) ELISA Kit |
EH4913 |
FN Test |
96T |
EUR 567.6 |
|
Description: Method of detection: Coated with Antigen, Competitive ELISA;Reacts with: Homo sapiens;Sensitivity: 18.75 ng/ml |
Mouse Bromodeoxyuridine(BrdU)ELISA Kit |
GA-E0163MS-48T |
GenAsia Biotech |
48T |
EUR 336 |
Mouse Bromodeoxyuridine(BrdU)ELISA Kit |
GA-E0163MS-96T |
GenAsia Biotech |
96T |
EUR 534 |
Rat Bromodeoxyuridine(BrdU)ELISA Kit |
GA-E0659RT-48T |
GenAsia Biotech |
48T |
EUR 317 |
Rat Bromodeoxyuridine(BrdU)ELISA Kit |
GA-E0659RT-96T |
GenAsia Biotech |
96T |
EUR 496 |
Human Bromodeoxyuridine(BrdU)ELISA Kit |
GA-E1920HM-48T |
GenAsia Biotech |
48T |
EUR 289 |
Human Bromodeoxyuridine(BrdU)ELISA Kit |
GA-E1920HM-96T |
GenAsia Biotech |
96T |
EUR 466 |
Guinea Pig BrDU ELISA Kit |
EGB0693 |
Abclonal |
96Tests |
EUR 521 |
BrdU Cell Proliferation Assay Kit |
C0300-050 |
GenDepot |
500 Assays |
EUR 928 |
BrdU Cell Proliferation Assay Kit |
C0300-100 |
GenDepot |
1000 Assays |
EUR 1312 |
BrdU Cell Proliferation Assay Kit |
C0300-500 |
GenDepot |
5000 Assays |
EUR 4438 |
Human Bromodeoxyuridine,BrdU ELISA Kit |
201-12-1904 |
SunredBio |
96 tests |
EUR 440 |
|
Description: A quantitative ELISA kit for measuring Human in samples from biological fluids. |
5-Bromo-2'-deoxyuridine (BrdU) |
40024 |
Biotium |
100MG |
EUR 109 |
Description: Minimum order quantity: 1 unit of 100MG |
CytoSelect BrdU Competitive ELISA Kit |
CBA-5098 |
Cell Biolabs |
96 assays |
EUR 543 |
A chemical modifier was mixed with the cIEF effluent in the flow-throw microvial to maintain the ESI stability and to mitigate ion suppression from the co-eluted carrier ampholytes and glycerol. Analysis of mAb samples have shown relative populations of two basic variants originating from C-terminal lysine process and acidic variant of deamidation.
The lysine clippings, deamidation, and sialic acid modification in oligosaccharide chains were revealed in infliximab. Two lysine clipping variants and a deamidated variant were observed in adalimumab. The duplicate analyses of a reference mAb demonstrated five charge variants separated by cIEF due to some unidentified modifications, as their mass spectra shared close similarities.
The mAb analyses demonstrated the feasibility of the cIEF-MS method, and they demonstrated how charge and structural variants and minor differences in therapeutic mAbs are observed with this technology. Online cIEF-MS is an information rich technology with high throughput, demonstrated by the initial data presented here.