Analysis of the Structure of 14 Therapeutic Antibodies Using Circular Dichroism Spectroscopy

Maria Bruque; Rodger Alison; Søren Vrønning Hoffmann; Nykola Jones; Jean Aucamp; Tim Dafforn; Owen Thomas

Analysis of the Structure of 14 Therapeutic Antibodies Using Circular Dichroism Spectroscopy

Maria Bruque, Rodger Alison, Søren Vrønning Hoffmann, Nykola Jones, Jean Aucamp, Tim Dafforn, Owen Thomas^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

Understanding the impact of manufacturing environment on therapeutic monoclonal antibody (mAb) structure requires new process analytical technology. Here we describe the creation of a new reference set for circular dichroism (CD) spectra of mAbs. Datasets of the highest quality were collected by synchrotron radiation CD for 14 different mAbs in both native and acid-stressed states. Deconvolution of far-UV spectra for the mAb cohort identified two current reference sets (SP175 and SMP180) as assigning accurate secondary structures irrespective of the analysis program employed. Scrutiny of spectra revealed signifi-cant variation in the far-UV and especially near-UV CD of the 14 mAbs. Two spectral features were found to be sensitive to changes in solution pH, i.e., the far-UV positive peak at 201-202 nm and near-UV negative exciton couplet around 230-240 nm. The latter feature offers attractive possibilities for in-line CD-based monitoring of mAb structure during manufacture.

Original language	English
Journal	Analytical Chemistry
Publication status	Accepted/In press - 27 Aug 2024

Bibliographical note

Not yet published as of 10/09/2024.

Cite this

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title = "Analysis of the Structure of 14 Therapeutic Antibodies Using Circular Dichroism Spectroscopy",

abstract = "Understanding the impact of manufacturing environment on therapeutic monoclonal antibody (mAb) structure requires new process analytical technology. Here we describe the creation of a new reference set for circular dichroism (CD) spectra of mAbs. Datasets of the highest quality were collected by synchrotron radiation CD for 14 different mAbs in both native and acid-stressed states. Deconvolution of far-UV spectra for the mAb cohort identified two current reference sets (SP175 and SMP180) as assigning accurate secondary structures irrespective of the analysis program employed. Scrutiny of spectra revealed signifi-cant variation in the far-UV and especially near-UV CD of the 14 mAbs. Two spectral features were found to be sensitive to changes in solution pH, i.e., the far-UV positive peak at 201-202 nm and near-UV negative exciton couplet around 230-240 nm. The latter feature offers attractive possibilities for in-line CD-based monitoring of mAb structure during manufacture.",

author = "Maria Bruque and Rodger Alison and {Vr{\o}nning Hoffmann}, S{\o}ren and Nykola Jones and Jean Aucamp and Tim Dafforn and Owen Thomas",

note = "Not yet published as of 10/09/2024.",

year = "2024",

month = aug,

day = "27",

language = "English",

journal = "Analytical Chemistry",

issn = "0003-2700",

publisher = "American Chemical Society",

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TY - JOUR

T1 - Analysis of the Structure of 14 Therapeutic Antibodies Using Circular Dichroism Spectroscopy

AU - Bruque, Maria

AU - Alison, Rodger

AU - Vrønning Hoffmann, Søren

AU - Jones, Nykola

AU - Aucamp, Jean

AU - Dafforn, Tim

AU - Thomas, Owen

N1 - Not yet published as of 10/09/2024.

PY - 2024/8/27

Y1 - 2024/8/27

N2 - Understanding the impact of manufacturing environment on therapeutic monoclonal antibody (mAb) structure requires new process analytical technology. Here we describe the creation of a new reference set for circular dichroism (CD) spectra of mAbs. Datasets of the highest quality were collected by synchrotron radiation CD for 14 different mAbs in both native and acid-stressed states. Deconvolution of far-UV spectra for the mAb cohort identified two current reference sets (SP175 and SMP180) as assigning accurate secondary structures irrespective of the analysis program employed. Scrutiny of spectra revealed signifi-cant variation in the far-UV and especially near-UV CD of the 14 mAbs. Two spectral features were found to be sensitive to changes in solution pH, i.e., the far-UV positive peak at 201-202 nm and near-UV negative exciton couplet around 230-240 nm. The latter feature offers attractive possibilities for in-line CD-based monitoring of mAb structure during manufacture.

AB - Understanding the impact of manufacturing environment on therapeutic monoclonal antibody (mAb) structure requires new process analytical technology. Here we describe the creation of a new reference set for circular dichroism (CD) spectra of mAbs. Datasets of the highest quality were collected by synchrotron radiation CD for 14 different mAbs in both native and acid-stressed states. Deconvolution of far-UV spectra for the mAb cohort identified two current reference sets (SP175 and SMP180) as assigning accurate secondary structures irrespective of the analysis program employed. Scrutiny of spectra revealed signifi-cant variation in the far-UV and especially near-UV CD of the 14 mAbs. Two spectral features were found to be sensitive to changes in solution pH, i.e., the far-UV positive peak at 201-202 nm and near-UV negative exciton couplet around 230-240 nm. The latter feature offers attractive possibilities for in-line CD-based monitoring of mAb structure during manufacture.

UR - http://pubs.acs.org/journal/iecac0

M3 - Article

SN - 0003-2700

JO - Analytical Chemistry

JF - Analytical Chemistry

ER -

Analysis of the Structure of 14 Therapeutic Antibodies Using Circular Dichroism Spectroscopy

Abstract

Bibliographical note

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