DIFFERENTIAL SCANNING CALORIMETRY (DSC)

Differential scanning calorimetry (DSC) is a thermoanalytical technique in which the difference in the amount of heat required to increase the temperature of a sample and reference is measured as a function of temperature. Endothermic samples will need more heat flux during analysis to increase the temperature at the same rate as the reference material. Otherwise, exothermic materials will require less heat flux to raise the temperature. The DSC curve indicates exothermic peaks upward and endothermic peaks downward.

Nano DSC for protein characterization

NANO DSC

  • The Nano DSC is designed to measure the amount of heat absorbed or released by dilute in-solution bio-molecules as they are heated or cooled. Macromolecules such as proteins respond to heating or cooling by unfolding at a characteristic temperature. 
  • The more intrinsically stable the biopolymer, the higher the midpoint temperature of the unfolding transition. As these processes often exchange microjoule levels of heat, the sensitivity of the Nano DSC is critical for successful investigation of the reaction.
  • The Nano DSC obtains data with less sample than competitive designs and produces unmatched short term noise (±15 nanowatts) and baseline reproducibility (±28 nanowatts). 
Nano DSC

Nano DSC Specifications

 

Short-term Noise 

0.015 µWatts

Baseline Stability

±0.028 µWatts

Response Time 

7 seconds

Operating Temperature

-10 °C to 130 °C or 160 °C

Temperature Scan Rate

0.05 °C to 2°C/minute

Cell Volume 

0.30 ml

Cell Geometry 

Fixed capillary

Cell Composition

Platinum

Heat Measurement Type 

Power Compensation

HIGHER ORDER STRUCTURE ANALYSIS BY NANO DSC

Protein maintains its higher order structure using multiple ionic, non-ionic, hydrophobic and other interactions between its subunits. However, at higher temperature when these interactions are disrupted protein unfolds and eventually denatures. 

Using Differential Scanning Calorimetry, that collects information on heat release when protein is unfolding, its melting temperature (a temperature where protein denatures) can be measured. Typically, melting temperature of a protein is its unique property and can be used as a comparative parameter to establish integrity of biotherapeutic products. 

HIGHER ORDER STRUCTURE ANALYSIS BY NANO DSC

NANO DSC – CASE STUDY

Property

Value

Name

210326_Bevacizumab.dsc / scan 1

User

analyst

Serial Number

K20342

Start Time

26-03-2021 13:31:59

Sample Name

Sample Name: 210326_bevacizumab

Concentration – 0.05 M sodium phosphate buffer pH 6.8

Scan Rate (°C/min)

1.0

Lower Temp 

(°C)

25.0

Upper Temp 

(°C)

100.0

Scan Blank

210326_blank.dsc / scan 1

NANO DSC – CASE STUDY

NANO DSC ANALYSIS AT CBA

 

Higher order Structure Analysis using NanoDSC

Sample requirement

Amount- 3-6 ml (1-2 mg/ml)

Formulation buffer- 10 ml

Deliverables

Thermogram of specific heat capacity Vs Temperature, apparent Tm

 

Information required from client

  • Concentration 
  • Molecular weight
  • Buffer composition (in case of formulation)

SERVICES OFFERED BY CBA

  • Protein Intact Mass Analysis using HRMS
  • Peptide Mapping using HRMS
  • Post Translational Modifications Analysis using HRMS
  • Host Cell Protein Impurity Identification using HRMS
  • Host Cell Protein Impurity Profiling using SDS-PAGE
  • Secondary Structure Analysis using far-UV CD Spectroscopy
  • Tertiary Structure Analysis using near-UV CD Spectroscopy
  • Tertiary Structure Analysis using Fluorescence Spectroscopy
  • Higher Order Structure Analysis using Fluorescence Spectroscopy
  • Secondary Structure Analysis using FTIR Spectroscopy
  • Higher Order Structure Analysis using DSC
  • Aggregate/Multimers Analysis using SEC-MALS
  • Protein Protein Interaction Analysis using SPR
  • Biosimilarity assessment Project

SERVICES OFFERED BY CBA

Dr. Smita Kale

PI- Center for BioPharma Analysis

Venture Center

100, NCL Innovation Park, Dr. Homi Bhabha Road, Pune – 08

Phone: 020-2586-5877 | Mobile: +91-9923147592

Email: smita@venturecenter.co.in

Mr. Yashodhan Shende

Business Development Executive

Venture Center

100, NCL Innovation Park, Dr. Homi Bhabha Road, Pune – 08

Phone: 020-2586-5877 | Mobile: +91-7755920096 

Email: yashodhan@venturecenter.co.in

For more information, please contact:

Website: https://bioanalysis.in/