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Open Dataset XSUJ5ICU


SEC-SAXS-MALS for mAb4 ( SinoBiological: 40143-R004) + mAb2 ( SinoBiological: 40143-MM08 ) + NPNTD

Abstract:

We conducted a structural characterization of mAbs against the N-terminal domain of nucleocapsid protein (NPNTD) from SARS-CoV-2 using small-angle X-ray scattering (SAXS). Our solution-based results distinguished the mAbs’ flexibility and how this flexibility impacts the assembly of multiple mAbs on an antigen. By pairing two mAbs that bind different epitopes on the NPNTD, we show that flexible mAbs form a closed sandwich-like complex. With rigid mAbs, a juxtaposition of the Fabs is prevented, enforcing a linear arrangement of the mAb pair, which facilitates further mAb polymerization.

Experimental description:

For SEC-MALS-SAXS experiments, 60 uL of a sample containing mAb2 ~1.5 mg/mL + mAb4 ~1.5 mg/ml + NPNTD in 1:5 molar ratio was prepared in PBS pH 7.4 buffer. SEC-MALS-SAXS was collected at the SIBYLS beamline (BL 12.3.1) at the Advanced Light Source (ALS) at Lawrence Berkeley National Laboratory (LBNL) in Berkeley, California.

File description:

*.sec > unsubtracted SAXS curves for 600 two-second exposures across the SEC elution profile ready for SCATTER analysis *.dat > final merged SAXS profile for main SEC peak containing mAb2/mAb4/NPNTD complex *pdf > MALS results

Data collection technique:

SEC-SAXS

Wavelength

Sample to Detector Distance


Created: 2021-03-05

Updated: 2021-05-21

Submitting Author:

Michal Hammel

Lawrence Berkeley National Laboratory, The SIBYLS Beamline

United States of America

mhamm[email protected]

Collaborators:

https://www.biorxiv.org/content/10.1101/2021.01.13.426597v1

Project Leader:

Michal Hammel

[email protected]


Data Download Terms:

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Complete Set of SAXS Data Files:

The complete SAXS dataset is downloadable as a zip file.


Sample SAXS Data Files (total 1):

These are individual SAXS data files that are included in the complete dataset, but are available separately for quick download as examples of what is contained in the zip file.


Supplemental Data and Supporting Materials (total 1):

These data and materials may include X-ray crystal structure coordinates, multi-angle light scattering data, .etc. It may also include additional details or methods pertaining to the SAXS experiments, or the researcher's interpretations of the results.

  • Covid_anti3[070920_Covid].pdf Download

Open SAXS data analysis sites in a new tab using these links:

SAXS Similarity SAXS FrameSlice


Samples:

  • Macromolecule 1: mAb2
    • Sample Full Name: mAb2 ( SinoBiological: 40143-MM08 )
    • Sample Type: Protein
    • Source Organism:
    • Source Organism NCBI Taxonomy ID:
    • Expression System:
    • Expression NCBI Taxonomy ID:
    • Sequence or Chemical Formula:
  • Macromolecule 2: mAb4
    • Sample Full Name: mAb4 ( SinoBiological: 40143-R004 )
    • Sample Type: Protein
    • Source Organism:
    • Source Organism NCBI Taxonomy ID:
    • Expression System:
    • Expression NCBI Taxonomy ID:
    • Sequence or Chemical Formula:
  • Macromolecule 3: NPNTD
    • Sample Full Name: N-terminal domain of nucleocapsid protein from SARS-CoV-2
    • Sample Type: Protein
    • Source Organism:
    • Source Organism NCBI Taxonomy ID:
    • Expression System:
    • Expression NCBI Taxonomy ID:
    • Sequence or Chemical Formula: