Key facts
- Brand
- Agilent
- Model
- 1200 Series HPLC
- Documents
- 4
- Languages
- EN
- Last added
- 2012-07-02
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Agilent Antibody Analysis by ESI-TOF LC/MS Application Note
Monoclonal antibodies (mAbs) are complex structures requiring accurate characterization, particularly for determining molecular weight. Traditional methods like gel analysis or MALDI-TOF often suffer from imprecision or lengthy processing times.
This study evaluates the combination of Agilent Technologies Poroshell column, 1200 HPLC system, and 6210 Time-of-Flight mass spectrometer for analyzing intact IgG4 antibodies. The method achieves a short retention time of 2.78 minutes with good chromatographic peak shape. Mass spectral analysis successfully resolved various antibody subpopulations, demonstrating routine mass accuracy better than 25 ppm.
Agilent 1200 Series HPLC specifications
| Retention time for intact IgG4 | 2.78 min |
|---|---|
| Routine mass accuracy | better than 25 ppm |
| Total analysis time | 9-minute |
| HPLC flow rate | ~500 l/min |
| Column oven temperature | 90°C |
| Capillary voltage (TOF MS) | 5500 V |
| Drying gas temperature | 300°C |
Source: Agilent Antibody Analysis by ESI-TOF LC/MS Application Note
All Agilent 1200 Series HPLC documents by type
How to use the product for one specific task, with worked examples.
- Agilent Antibody Analysis by ESI-TOF LC/MS Application Note 12 p
- Agilent 1200 Series HPLC/UV Melamine Determination Application Note 6 p
- Agilent 1200 Series HPLC System Application Note: Analysis of FAME and TG in Biodiesel 4 p
- Agilent Low-level Speciated Analysis of Cr(III) and Cr(VI) Using LC(IC)-ICP-MS Application Note 6 p
Questions answered in these manuals
What is the routine mass accuracy demonstrated by the system?
The routine mass accuracy demonstrated by the TOF MS was better than 25 ppm.
From: Agilent Antibody Analysis by ESI-TOF LC/MS Application Note
Which column is used for the optimal chromatographic peak shape?
The Poroshell C8 column was used, resulting in very good chromatographic peak shape.
From: Agilent Antibody Analysis by ESI-TOF LC/MS Application Note
What is the limit of detection for melamine in this method?
The limit of detection is 10 pg/kg.
From: Agilent 1200 Series HPLC/UV Melamine Determination Application Note
What is the linear calibration range for melamine?
Linear calibration curves were obtained over the range of 1 to 20 mg/kg.
From: Agilent 1200 Series HPLC/UV Melamine Determination Application Note
Which detector is used for this analysis?
A refractive index detector (RID) is used.
From: Agilent 1200 Series HPLC System Application Note: Analysis of FAME and TG in Biodiesel
What is the quantification limit for triglyceride analysis using this system?
The system can detect small amounts of triglyceride with a quantification limit of 1 g/10 pL.
From: Agilent 1200 Series HPLC System Application Note: Analysis of FAME and TG in Biodiesel