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TitleIncreasing sensitivity of antibody-antigen interactions using photo-cross-linking.
Journal, issue, pagesCell Rep Methods, Vol. 3, Issue 6, Page 100509, Year 2023
Publish dateJun 26, 2023
AuthorsAlba Torrents de la Peña / Leigh M Sewall / Rebeca de Paiva Froes Rocha / Abigail M Jackson / Payal P Pratap / Sandhya Bangaru / Christopher A Cottrell / Subhasis Mohanty / Albert C Shaw / Andrew B Ward /
PubMed AbstractUnderstanding antibody-antigen interactions in a polyclonal immune response in humans and animal models is critical for rational vaccine design. Current approaches typically characterize antibodies ...Understanding antibody-antigen interactions in a polyclonal immune response in humans and animal models is critical for rational vaccine design. Current approaches typically characterize antibodies that are functionally relevant or highly abundant. Here, we use photo-cross-linking and single-particle electron microscopy to increase antibody detection and unveil epitopes of low-affinity and low-abundance antibodies, leading to a broader structural characterization of polyclonal immune responses. We employed this approach across three different viral glycoproteins and showed increased sensitivity of detection relative to currently used methods. Results were most noticeable in early and late time points of a polyclonal immune response. Additionally, the use of photo-cross-linking revealed intermediate antibody binding states and demonstrated a distinctive way to study antibody binding mechanisms. This technique can be used to structurally characterize the landscape of a polyclonal immune response of patients in vaccination or post-infection studies at early time points, allowing for rapid iterative design of vaccine immunogens.
External linksCell Rep Methods / PubMed:37426749 / PubMed Central
MethodsEM (single particle)
Resolution25.0 Å
Structure data

EMDB-27346: nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab from individual 182419 at day 0
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27347: nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182419 at day 0 using Sulfo-SDAD photo-cross-linker
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27348: nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182419 at day 2
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27349: nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182419 at day 2 using Sulfo-SDAD photo-cross-linker
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27350: nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182419 at day 7 using Sulfo-SDAD photo-cross-linker
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27351: nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182419 at day 28
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27352: nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182419 at day 28 using Sulfo-SDAD photo-cross-linker
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27353: nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182419 at day 70
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27354: nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182419 at day 70 using Sulfo-SDAD photo-cross-linker
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27355: nsEM map of hemagglutinin H3 A/Sing/INFIMH/16 complexed with polyclonal Fab samples from individual 182419 at day 0
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27356: nsEM map of hemagglutinin H3 A/Sing/INFIMH/16 complexed with polyclonal Fab samples from individual 182419 at day 0 using Sulfo-SDAD photo-cross-linker
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27357: nsEM map of hemagglutinin H3 A/Sing/INFIMH/16 complexed with polyclonal Fab samples from individual 182419 at day 2
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27358: nsEM map of hemagglutinin H3 A/Sing/INFIMH/16 complexed with polyclonal Fab samples from individual 182419 at day 2 using Sulfo-SDAD photo-cross-linker
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27359: nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182419 at day 7
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27360: nsEM map of hemagglutinin H3 A/Sing/INFIMH/16 complexed with polyclonal Fab samples from individual 182419 at day 7
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27361: nsEM map of hemagglutinin H3 A/Sing/INFIMH/16 complexed with polyclonal Fab samples from individual 182419 at day 7 using Sulfo-SDAD photo-cross-linker
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27362: nsEM map of hemagglutinin H3 A/Sing/INFIMH/16 complexed with polyclonal Fab samples from individual 182419 at day 28
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27363: nsEM map of hemagglutinin H3 A/Sing/INFIMH/16 complexed with polyclonal Fab samples from individual 182419 at day 28 using Sulfo-SDAD photo-cross-linker
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27364: nsEM map of hemagglutinin H3 A/Sing/INFIMH/16 complexed with polyclonal Fab samples from individual 182419 at day 70
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27365: nsEM map of hemagglutinin H3 A/Sing/INFIMH/16 complexed with polyclonal Fab samples from individual 182419 at day 70 using Sulfo-SDAD photo-cross-linker
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27366: nsEM map of beta coronavirus OC43 spike complexed with polyclonal Fab samples from individual 182419 at day 0
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27367: nsEM map of beta coronavirus OC43 spike complexed with polyclonal Fab samples from individual 182419 at day 0 using Sulfo-SDAD photo-cross-linker
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27368: nsEM map of beta coronavirus HKU1 spike complexed with polyclonal Fab samples from individual 182419 at day 0
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27369: nsEM map of beta coronavirus HKU1 spike complexed with polyclonal Fab samples from individual 182419 at day 0 using Sulfo-SDAD photo-cross-linker
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27370: nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182322 at day 28
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27371: nsEM map of hemagglutinin H1 A/Mich/045/15 complexed with polyclonal Fab samples from individual 182322 at day 28 using Sulfo-SDAD photo-cross-linker
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27372: nsEM map of hemagglutinin H3 A/Sing/INFIMH/16 complexed with polyclonal Fab samples from individual 182322 at day 28
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27373: nsEM map of hemagglutinin H3 A/Sing/INFIMH/16 complexed with polyclonal Fab samples from individual 182322 at day 28 using Sulfo-SDAD photo-cross-linker
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27374: nsEM map of beta coronavirus OC43 spike complexed with polyclonal Fab samples from individual 182322 at day 0
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27375: nsEM map of beta coronavirus OC43 spike complexed with polyclonal Fab samples from individual 182322 at day 0 using Sulfo-SDAD photo-cross-linker
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27376: nsEM map of beta coronavirus HKU1 spike complexed with polyclonal Fab samples from individual 182322 at day 0
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27377: nsEM map of beta coronavirus HKU1 spike complexed with polyclonal Fab samples from individual 182322 at day 0 using Sulfo-SDAD crosslinker
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27378: nsEM map of influenza hemagglutinin HA A/Mich/045/15 bound to H5.28 antibody
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27379: nsEM map of influenza HA A/Mich/045/15 photo-cross-linked trimer using Sulfo-SDAD crosslinker
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27380: nsEM map of HIV soluble BG505 SOSIP glycoprotein bound to 3E1 using Sulfo-SDAD photo-cross-linker
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-27381: nsEM map of HIV BG505 glycoprotein complexed with polyclonal Fab samples from rhesus macaque rT8640 at week 18 using Sulfo-SDAD photo-cross-linker
Method: EM (single particle) / Resolution: 25.0 Å

Source
  • Homo sapiens (human)

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