Page 34 - Pharmaceutical- Guide to Biopharmaceutical

Characterization Quality Control

Quantitation of Proteins BioSpec-nano

Quantitation of Proteins Using benefits

BioSpec-nano Cell Line Optimization
• Measure the concentration of proteins or check the purity.
• Measure sample quantities as small as 1 μL.
Operating Principle and Features
• Achieve low carryover with the automatic wiping function.
The BioSpec-nano has two available optical path lengths, 0.2 mm and
0.7 mm, and can quantify proteins and nucleic acids in sample quantities
as low as 1 µL. Samples can also be measured using an optional cell with Culture
a 5 mm optical path length (for 2 mL sample quantities). That means
only a small sample quantity is needed for quick protein concentration
measurements or to check spectra.
A wiping mechanism enables automatic cleaning between samples,
ensuring extremely low carryover and reducing inconsistencies that
may occur with manual cleaning (Fig. 1).

Wipe

Fig. 3 Analytical Condition Selection Window Purification
Results

Measurement results are shown in Fig. 4. The results show a
concentration of approx. 1000 μg/mL, which is the same as prepared.
Target Sample concentration values and spectra can be confirmed in the detail
display window. Results can be output in either CSV or PDF format.
Fig. 1 Automatic Wiping Function
Measurement Method Characterization

Measurement samples were prepared by diluting bovine immunoglobulin
(IgG) to 1000 μg/mL with phosphate buffered saline. With the optical Specifications
path length set to 0.7 mm (Fig. 2), 4 μL of the sample was dripped onto
the target to measure the optical density (OD) at 280 nm. The protein Instrument BioSpec-nano
concentration is calculated based on the molar absorption coefficient (ε
= 280) and molecular weight values entered in the software's analytical Wavelength range 220 to 800 nm
condition selection window (Fig. 3). The 280 ε value can also be Spectrum bandwidth 3 nm
calculated by the software if the number of tryptophan, tyrosine, and
cysteine residues in the amino acid sequence is entered. Wavelength accuracy ±1 nm Quality Control
Pathlength 0.2 mm, 0.7 mm
4.6
Photometric value unit OD (Optical Density), absorbance converted with 10 mm pathlength
The optical path length can be 3.45 Sample volume 1 µL min. (pathlength: 0.2 mm)
2 µL min. (pathlength: 0.7 mm)
selected by simply moving the lever.
Light source Xenon flash lamp
2.3
Monochromator Holographic grating
Detector Photo diode array
1.15
Auto wiping function Provided Pharmacokinetics
Spectrum measuring time 3 sec
0
Fig. 2 Optical Path Length Setting Area
240 250 260 270 280 290 300 310 320 Quantitation range Pathlength 0.2 mm, 1 to 75 OD, 50 to 3,700 ng/µL
Conclusion Wavelength (nm) Pathlength 0.7 mm, 0.3 to 21 OD, 15 to 1,000 ng/µL
Fig. 4 Analytical Results for 0.7 mm Optical Path Length Optional 5 mm pathlength cell, 0.04 to 3 OD, 2 to 150 ng/μL
Using the BioSpec-nano, sample concentrations can be measured from Dimensions W 210 mm x D 214 mm x H 417 mm
small sample quantities of 1 to 4 μL by simply dripping the sample onto Application Examples
the stage. That eliminates any need for any manual processes, such as Weight 7 kg Others
raising/lowering an arm, placing a cell in position, or wiping off the sample • Measuring single- or double-stranded DNA concentrations (refer to p. 6) Analysis mode Simple nucleic acid quantitation, labeled nucleic acid quantitation, protein quantitation,
after each measurement. In addition to concentration determination, OD • Measuring labeled nucleic acid concentrations labeled protein quantitation, photometric measurement
values can be displayed for any user-specified wavelength. • Measuring RNA concentrations Note: The droplet formation status will affect analysis results. Measure quantities that are large enough to enable proper droplet formation.

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