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Structure paper

TitleFrom structure to sequence: Antibody discovery using cryoEM.
Journal, issue, pagesSci Adv, Vol. 8, Issue 3, Page eabk2039, Year 2022
Publish dateJan 21, 2022
AuthorsAleksandar Antanasijevic / Charles A Bowman / Robert N Kirchdoerfer / Christopher A Cottrell / Gabriel Ozorowski / Amit A Upadhyay / Kimberly M Cirelli / Diane G Carnathan / Chiamaka A Enemuo / Leigh M Sewall / Bartek Nogal / Fangzhu Zhao / Bettina Groschel / William R Schief / Devin Sok / Guido Silvestri / Shane Crotty / Steven E Bosinger / Andrew B Ward /
PubMed AbstractOne of the rate-limiting steps in analyzing immune responses to vaccines or infections is the isolation and characterization of monoclonal antibodies. Here, we present a hybrid structural and ...One of the rate-limiting steps in analyzing immune responses to vaccines or infections is the isolation and characterization of monoclonal antibodies. Here, we present a hybrid structural and bioinformatic approach to directly assign the heavy and light chains, identify complementarity-determining regions, and discover sequences from cryoEM density maps of serum-derived polyclonal antibodies bound to an antigen. When combined with next-generation sequencing of immune repertoires, we were able to specifically identify clonal family members, synthesize the monoclonal antibodies, and confirm that they interact with the antigen in a manner equivalent to the corresponding polyclonal antibodies. This structure-based approach for identification of monoclonal antibodies from polyclonal sera opens new avenues for analysis of immune responses and iterative vaccine design.
External linksSci Adv / PubMed:35044813 / PubMed Central
MethodsEM (single particle)
Resolution3.3 - 15.9 Å
Structure data

EMDB-23778:
Negative stain EM map of BG505 SOSIP in complex with Rh4O9.8 monoclonal antibody (as Fab fragment)
Method: EM (single particle) / Resolution: 15.9 Å

23779

7mdt

EMDB-23779: BG505 SOSIP.v5.2 in complex with the polyclonal Fab pAbC-1 from animal Rh.4O9
PDB-7mdt: BG505 SOSIP.v5.2 in complex with the monoclonal antibody Rh4O9.8 (as Fab fragment)
Method: EM (single particle) / Resolution: 3.6 Å

23780

7mdu

EMDB-23780, PDB-7mdu:
BG505 SOSIP MD39 in complex with the monoclonal antibodies Rh.33104 mAb.1 and RM20A3
Method: EM (single particle) / Resolution: 3.3 Å

23801

7mep

EMDB-23801, PDB-7mep:
BG505 SOSIP.v5.2(7S) in complex with the monoclonal antibodies Rh.33172 mAb.1 and RM19R
Method: EM (single particle) / Resolution: 3.5 Å

Chemicals

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose / N-Acetylglucosamine

Source
  • human immunodeficiency virus
  • human immunodeficiency virus 1
  • macaca mulatta (Rhesus monkey)
KeywordsVIRAL PROTEIN / monoclonal antibody / polyclonal antibody / immune complex / monoclonal antibodies / HIV-1 / BG505 SOSIP

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