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[AQS3®pro] 단백질 2차 구조(Secondary Protein Structure) 특성 분석 어플리케이션
인성크로마텍(주)
Date : 2021.08.04
분류 : Analytical Products > RedShiftBio


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Application Notes & White Papers.PNG

 

 

"하기의 App Notes 중 필요하신 자료를 문의 주시면 이메일로 송부 드리겠습니다"

 

 

 

Biosimilar Structural Comparison of Commercially Sourced Reference Standards by MMS Rapidly 

Detects Subtle but Critical Differences to Correctly Predict Activity for Use in an ELISA Product 

 

July 29, 2021

 

 

 

This Application Note Includes:


• This study confidently detected <2% structural difference between samples to correctly predict activity amongst the

  commercially sourced materials.

• MMS successfully identifies the 5 of 13 BSA biosimilar samples that retained acceptable activity upon inclusion in
  the ELISA product through structural comparison to a reference of known activity. 

 

It was determined that many of the Bovine Serum Albumin (BSA) reference candidates did not exhibit acceptable activity after

inclusion into the final product though all purchased samples shared the same name and identification. The inconsistency in 

activity causes delays, increases cost of manufacture, and adds an element of variability. Current in-house biophysical 

techniques fail to correctly identify the inactive candidates prior to product inclusion due to inconclusive results or a lack of 

sensitivity.Thus a more accurate, reliable analytical method was sought using MMS. The AQS3pro was able to provide a more 

detailed analysis and measurements through a simple numerical output reporting the percent similarity of each sample’s 

spectra relative to the active in-house standard reference was sufficient to correctly identify the active 5 candidates.

 

 

 

 

 

Biosimilar Comparison and Accelerated Stability Predictions Based on <2% Secondary Structure 

Differences Using Microfluidic Modulation Spectroscopy (MMS)

 

June 24, 2021

 

 

This Application Note Includes:

 

 • Microfluidic Modulation Spectroscopy (MMS) as a tool that measures differences in secondary structure of <2% between

   samples due to highly reproducible replicate measurements generated by the AQS3pro.

 • The power of MMS to predict the relative order of sample stability based on very small secondary structure differences

   detected between sample pairs of biosimilar molecules incubated at 4°C and 30°C for 8 weeks. 

 

Biosimilar Comparison and Accelerated Stability Predictions Based on <2% Secondary Structure Differences Using Microfluidic

Modulation Spectroscopy (MMS) Biosimilar structure analysis and stability predictions require instrumentation sensitive

enough to detect critical small changes in secondary structure. The ability to see these structural differences between

engineered biosimilars and their innovator molecules is vital to ensure successful drug approval. Biosimilar structure analysis

and stability predictions …“Biosimilar Comparison and Accelerated Stability Predictions Based on <2% Secondary Structure 

Differences Using Microfluidic Modulation Spectroscopy (MMS)"


 

 

 

 

Detection of Pressure-Induced Protein Aggregation Using 

Microfluidic Modulation Spectroscopy (MMS)

 

April 8, 2021

 

 

The ability to detect protein aggregation is important at all stages of drug development. Early detection of aggregation is

most desirable to inform development decisions since aggregation can negatively affect the functionality of a protein.

 

This Application Note Includes:

 

• Highlights the significant changes present in the spectral regions of 1624 and 1640cm-1 in the Second Derivative plot that

  can be seen only subtly in the Absolute Absorbance spectral plot.

• Weighted Spectral Difference (WSD) as a metric used to monitor change in sample secondary structure for this aggregation

  study

• Strong linear correlation between increasing amounts of percent aggregation and increasing amounts of measured

  antiparallel beta-sheet content

 

Pressure-Induced Protein Aggregation Detection Protein aggregation is a recognized signal of instability and can lead to the

loss of protein function. It is therefore crucial to detect protein aggregation early in the drug development process to inform

further drug development decisions. Pressure, a stressor used for generating aggregates by impacting noncovalent interactions

without the need to change temperature or solvents1, was employed to create aggregated protein for this spiked study.

 

 

 

 

 

Secondary Structure Analysis: Using Microfluidic Modulation Spectroscopy to Determine 

the Structural Effect of pH on the Peptide Hormone Insulin

 

December 1, 2020

 

 

This Application Note Includes:

 

• Determines the structural effects of low pH on insulin using MMS

• Measures protein secondary structure by combining infrared spectroscopy with microfluidics to enhance sensitivity and accuracy

• Achieves higher sensitivity across a wider concentration range compared to current FTIR techniques

 

Protein Analysis Tools: Characterizing Secondary Structure Analysis Of Insulin

 

Secondary structure characterization is an essential part of drug development, however, the tools for measuring secondary

structure of small proteins and peptides are limited. Microfluidic Modulation Spectroscopy (MMS), a new characterization

platform that powers the AQS3pro system developed by RedshiftBio, measures protein secondary structure by combining infrared

spectroscopy with microfluidics to enhance sensitivity and accuracy.

 

 

 

 

 

Detecting Protein Conformational Change Due to Ligand Binding and Stabilization Using MMS

 

December 1, 2020

 

 

This Application Note Highlights:

 

• Protein was tested with two different ligands: one that completely stabilizes the protein against heat stress, and the other that

  moderately stabilizes it against heat stress.

• Determining of the secondary structure of proteins with and without ligands, offeringa more complete understanding of how

  the ligand alters the protein’s function

• Modulating between sample and reference buffer with the flow cell utilized byThe AQS3pro

• Enabling realtime, highly accurate and compatable buffer subtraction with MMS,for use with any ligand.

 

Detecting Protein Conformational Change

 

Ligand binding can affect the function of proteins and often causes protein conformational change. Since form fits function, 

determining the secondary structure of proteins with and without ligands is essential for a more complete understanding of 

how the ligand alters the protein’s function. RedShiftBio has developed a new technology called Microfluidic Modulation 

Spectroscopy (MMS) that measures protein secondary structure by combining infrared spectroscopy with microfluidics to 

enhance sensitivity and accuracy.

 

 

 

 

 

Assessment of Quantitation Linearity for Bovine Serum Albumin Using MMS vs FTIR

 

December 1, 2020

 

 

This Application Note Includes:

 

• BSA was prepared as a series and examined by MMS to measure the concentration linearity of the series.

• Measures protein secondary structure by combining infrared spectroscopy with microfluidics to enhance sensitivity and accuracy

• Achieves higher sensitivity across a wider concentration range compared to current FTIR techniques

 

FTIR Protein Analysis Of MMS

 

Microfluidic Modulation Spectroscopy (MMS) is a revolutionary new technology capable of highly sensitive and reproducible

measurements of protein secondary structure using IR absorbance in the Amide I band (1700-1600 cm-1). MMS is performed on

the AQS3pro system from RedShift BioAnalytics, and it generates measurements over a wide range of complex backgrounds. The

AQS3pro operates using a quantum cascade laser in combination with a microfluidic flow cell resulting in a wide dynamic range 

of 0.1 to >200 mg/mL for structural characterization, and further extended to 0.01 to >200 mg/mL for quantitation measurements.

 

 

 

 

 

Secondary Structure Analysis of NIST Monoclonal Antibody Reference Material 8671 

by Microfluidic Modulation Spectroscopy

 

October 1, 2020 

 

 

This Application Note Includes:

 

• NIST mAb RM 8671 is evaluated by MMS, which proves to be a uniquely valuable method for the measurement of higher order

  structure (HOS) across a wide concentration range

• MMS yields high quality, reproducible spectra over the measured concentration range.

• MMS achieves higher sensitivity across a wider concentration range comparedto current FTIR techniques

 

Characterizing Monoclonal Antibody Structure In 2016 the National Institute of Standards and Technology (NIST) made

NISTmAb Reference Material (RM) 8671 commercially available for the evaluation of methods used to characterize the structure of

monoclonal antibodies. This then serves as a performance control against which novel processes and emerging characterization 

technologies could be compared. Because this mAb is so widely characterized, its evaluation using Microfluidic Modulation 

Spectroscopy (MMS) is an important measurement and the first step in characterizing mAbs against a reputable standard.

 

 

 

 

 

Secondary Structure Determination using Microfluidic Modulation Spectroscopy 

in the Presence of DMSO

 

October 1, 2020

 

 

This Application Note Includes:

 

• lysozyme was dissolved in a buffer containing 0, 5, and 10% DMSO

• MMS generates an output that is 100 times brighter than traditional FTIR light sourcesby utilizing the ASQS3pro's Quantum 

  Casade Laser as the IR light source

• MMS can successfully be used for secondary structure determination in the presence of DMSO

• MMS results in a much greater signal-to-noise ratio and high sensitivity for accurate measurement of proteins ranging from

  0.1 to over 200 mg/mL

 

Secondary Structure Determination In The Presence Of DMSO

 

Characterizing biologic drug products is crucial in drug development and the presence of organic solvents and optically active

excipients typically hinders traditional characterization techniques. RedshiftBio has developed the AQS3pro, a novel 

characterization system powered by Microfluidic Modulation Spectroscopy (MMS) as a solution to overcoming these obstacles.

MMS combines Infrared (IR) spectroscopy with automatic sample handling and a micro-flow cell for accurate measurements with

no buffer interference. The system uses rapid modulation between a sample and its reference buffer to generate a differential 

spectrum for the most accurate buffer subtraction.This app note demonstrates that MMS can be used as an alternative to 

traditional secondary structure determination tools, when organic solvents and optically active excipients are present.

 

 

 

 

 

Monoclonal Antibody Analysis by Microfluidic Modulation Spectroscopy 

in a Complex Formulation Buffer (1 to 150 mg/mL)

 

March 1, 2019

 

 

This Application Note Includes:

 

• A native monoclonal antibody was measured by MMS in a complex buffer formulation at concentrations ranging from 

  1 to 150 mg/mL

• The use of simpler detectors without the need for liquid nitrogen cooling

• Demonstration of how MMS automatically introduces the sample protein solution and a matching reference buffer stream

  into a microfluidic flow cell

• How MMS modulates two fluids rapidly across theQuantum Cascade Laser beam path,producing nearly drift-free background

  compensated measurements

 

Monoclonal Antibodies Secondary Structure Determination

 

Microfluidic Modulation Spectroscopy (MMS) is a powerful infrared spectroscopy tool for protein structural analysis developed by

RedShift BioAnalytics. This technology provides significant increases in sensitivity, dynamic range, and accuracy for determination

of protein secondary structure relative to conventional mid-IR and far-UV CD techniques. This application note demonstrates the 

utility of the technique for the determination of monoclonal antibodies secondary structure at high buffer concentrations.

 

 

 

 

 

Thermal Denaturation Analysis of Bovine Serum Albumin over Wide Concentration Range 

by Microfluidic Modulation Spectroscopy

 

January 1, 2019

 

 

This Application Note Includes:

 

• In this App, MMS was used to study the heat-induced thermal denaturation of BSA samples

• Allows the use of simpler detectors without the need for liquid nitrogen cooling

• Differentiates small changes in secondary structure across a wide concentration range without the interference of excipients or

  loss of linearity at the lowest and highest concentrations

• Delivers information about aggregation, stability, HOS, and similarity in addition to quantitation results

 

Secondary Structure Analysis Tools For Measuring Thermal Denaturation

 

Microfluidic Modulation Spectroscopy (MMS) is a revolutionary new technology capable of highly sensitive and reproducible

measurements of protein secondary structure using IR absorbance in the Amide I band (1700-1600 cm-1). MMS is performed on

the AQS3pro system from RedShift BioAnalytics, and it generates measurements over a wide range of complex backgrounds,

including buffers that contain absorbing materials and scattering components such as reducing agents, adjuvants, surfactants,

and other excipients.

 

 

 

 

 

Microfluidic Modulation Spectroscopy (MMS) Fills an Analytical Gap with a Lower LOQ for 

Measuring Protein Misfolds and Structural Similarity

 

January 1, 2018

 

 

This Application Note Includes:

 

• MMS was used to study the heat-induced thermal denaturation of BSA samples

• Allows the use of simpler detectors without the need for liquid nitrogen cooling

• Differentiates small changes in secondary structure across a wide concentration range without the interference of excipients or

  loss of linearity at the lowest and highest concentrations

• Delivers information about aggregation, stability, HOS, and similarity in addition to quantitation results

 

Using MMS For Measuring Protein Misfolding And Aggregation

 

Protein misfolding in secondary structure can occur during all phases of drug development. A protein misfold represents a 

structural impurity and at any level can result in changed efficacy and increase the potential for immunogenicity. Improving 

spectroscopic methods for measuring low level impurities in secondary structure is necessary to maintain confidence in a 

protein’s integrity during all phases of drug development. Common structural characterization methods such as FTIR and CD 

have known limitations in reproducibility and sensitivity which adversely increase the lowest level of quantitation (LOQ) 

achievable when measuring structural impurities and similarity. Microfluidic Modulation Spectroscopy is a protein 

characterization method which generates reproducible high resolution measurements.

 

 

 

 

 

MMS For Protein Therapeutic Drug Analysis – White Paper

 

January 1, 2017

 

 

This Application Note Includes:

 

• In this App, RedShitBio provides an overview of the AQS3pro while it was still in late stages of development

• Simplifies measurement workflow

• Extends concentration range

• Measures secondary structure and fingerprinting with increased accuracy

• Demonstrates significant increases in sensitivity, dynamic range, and utility for determination of protein similarity 

  (fingerprinting), quantitation, protein secondary structure, and protein stability and aggregation through thermal and chemical

  denaturation methods with microfluidics to enhance sensitivity and accuracy

• Achieves higher sensitivity across a wider concentration range when measuring both chemical and thermal stability, 

  in addition to aggregation

 

Application Note Introduction

 

RedShiftBio’s MMS analyzer combines laser spectroscopy, microfluidics, and signal processing to provide a powerful, efficient 

tool that uses mid-infrared to directly probe the protein backbone and provide information on higher-order structure, aggregate

formation, protein stability, and concentration. Unlike other techniques, proteins can be measured over a broad concentration 

range with relative ease in a variety of environments without the issues of background fluorescence and light scattering, and 

complications related to protein size. 

 

 

 

 

 


 

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