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Epitope Mapping of Anti-mouse ACKR4 Monoclonal Antibodies Developed by N-Terminal Peptide Immunization.

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Τίτλος: Epitope Mapping of Anti-mouse ACKR4 Monoclonal Antibodies Developed by N-Terminal Peptide Immunization.
Συγγραφείς: Hirose M; Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan., Tanaka T; Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan., Yanaka M; Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan., Nakamura T; Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan., Kaneko MK; Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan., Suzuki H; Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan., Kato Y; Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.
Πηγή: Monoclonal antibodies in immunodiagnosis and immunotherapy [Monoclon Antib Immunodiagn Immunother] 2026 Jul-Aug; Vol. 45 (4), pp. 69-75. Date of Electronic Publication: 2026 Jun 16.
Τύπος έκδοσης: Journal Article
Γλώσσα: English
Στοιχεία περιοδικού: Publisher: Mary Ann Liebert, Inc Country of Publication: United States NLM ID: 101590955 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2167-9436 (Electronic) Linking ISSN: 21679436 NLM ISO Abbreviation: Monoclon Antib Immunodiagn Immunother Subsets: MEDLINE
Imprint Name(s): Original Publication: New Rochelle, NY : Mary Ann Liebert, Inc.
Ιατρικοί όροι (MeSH): Epitope Mapping*/methods , Antibodies, Monoclonal*/immunology , Epitopes*/immunology , Peptides*/immunology , Receptors, CXCR*/immunology , Receptors, CXCR*/genetics, Animals ; Mice ; Immunization ; Flow Cytometry ; Humans
Περίληψη: Leukocyte migration is a fundamental process in both innate and adaptive immune responses. This process is tightly regulated by chemokines and their cognate receptors. The bioavailability of chemokines is further modulated by atypical chemokine receptors (ACKRs), a subset of chemokine receptor-like molecules that lack coupling to canonical G protein-mediated signaling pathways. Among these, ACKR4 regulates dendritic cell migration through ligand scavenging and has been implicated in tumor progression in murine models. We previously established anti-mouse ACKR4 (mACKR4) mAbs, A4Mab-1, A4Mab-2, and A4Mab-3, by N-terminal peptide immunization. This study examined the binding epitopes of A4Mabs. Alanine (or glycine) scanning within the N-terminal region (amino acids 2-19) was performed using flow cytometry and Western blotting. Results showed that Tyr12 is required for recognition by A4Mab-1 in flow cytometry, whereas Tyr11, Tyr12, Glu14, Glu15, and Glu17 are required in Western blotting. For A4Mab-2, Tyr12, Glu15, and Asn16 are required in flow cytometry, whereas Tyr11, Tyr12, Tyr13, Glu15, and Asn16 are required in Western blotting. Additionally, Glu14, Asn16, and Glu17 are required for recognition by A4Mab-3 in flow cytometry. These findings contribute to the understanding of mACKR4 recognition by A4Mabs.
Competing Interests: Author Disclosure StatementThe authors have no conflicts of interest.
Contributed Indexing: Keywords: alanine scanning; epitope mapping; flow cytometry; monoclonal antibody; mouse ACKR4
Substance Nomenclature: 0 (Antibodies, Monoclonal)
0 (Epitopes)
0 (Peptides)
0 (Receptors, CXCR)
Entry Date(s): Date Created: 20260616 Date Completed: 20260702 Latest Revision: 20260702
Update Code: 20260703
DOI: 10.1177/21679436261461702
PMID: 42300976
Βάση Δεδομένων: MEDLINE
FullText Text:
  Availability: 0
Header DbId: cmedm
DbLabel: MEDLINE
An: 42300976
AccessLevel: 3
PubType: Academic Journal
PubTypeId: academicJournal
PreciseRelevancyScore: 0
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  Data: Epitope Mapping of Anti-mouse ACKR4 Monoclonal Antibodies Developed by N-Terminal Peptide Immunization.
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  Data: <searchLink fieldCode="AU" term="%22Hirose+M%22">Hirose M</searchLink>; Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.<br /><searchLink fieldCode="AU" term="%22Tanaka+T%22">Tanaka T</searchLink>; Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.<br /><searchLink fieldCode="AU" term="%22Yanaka+M%22">Yanaka M</searchLink>; Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.<br /><searchLink fieldCode="AU" term="%22Nakamura+T%22">Nakamura T</searchLink>; Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.<br /><searchLink fieldCode="AU" term="%22Kaneko+MK%22">Kaneko MK</searchLink>; Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.<br /><searchLink fieldCode="AU" term="%22Suzuki+H%22">Suzuki H</searchLink>; Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.<br /><searchLink fieldCode="AU" term="%22Kato+Y%22">Kato Y</searchLink>; Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.
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  Data: <searchLink fieldCode="JN" term="%22101590955%22">Monoclonal antibodies in immunodiagnosis and immunotherapy</searchLink> [Monoclon Antib Immunodiagn Immunother] 2026 Jul-Aug; Vol. 45 (4), pp. 69-75. <i>Date of Electronic Publication: </i>2026 Jun 16.
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  Data: Journal Article
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  Data: English
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  Data: <i>Publisher: </i><searchLink fieldCode="PB" term="%22Mary+Ann+Liebert%2C+Inc%22">Mary Ann Liebert, Inc </searchLink><i>Country of Publication: </i>United States <i>NLM ID: </i>101590955 <i>Publication Model: </i>Print-Electronic <i>Cited Medium: </i>Internet <i>ISSN: </i>2167-9436 (Electronic) <i>Linking ISSN: </i><searchLink fieldCode="IS" term="%2221679436%22">21679436 </searchLink><i>NLM ISO Abbreviation: </i>Monoclon Antib Immunodiagn Immunother <i>Subsets: </i>MEDLINE
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  Data: <i>Original Publication</i>: New Rochelle, NY : Mary Ann Liebert, Inc.
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  Label: MeSH Terms
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  Data: <searchLink fieldCode="MM" term="%22Epitope+Mapping%22">Epitope Mapping*</searchLink>/<searchLink fieldCode="MM" term="%22Epitope+Mapping+methods%22">methods</searchLink> <br /><searchLink fieldCode="MM" term="%22Antibodies%2C+Monoclonal%22">Antibodies, Monoclonal*</searchLink>/<searchLink fieldCode="MM" term="%22Antibodies%2C+Monoclonal+immunology%22">immunology</searchLink> <br /><searchLink fieldCode="MM" term="%22Epitopes%22">Epitopes*</searchLink>/<searchLink fieldCode="MM" term="%22Epitopes+immunology%22">immunology</searchLink> <br /><searchLink fieldCode="MM" term="%22Peptides%22">Peptides*</searchLink>/<searchLink fieldCode="MM" term="%22Peptides+immunology%22">immunology</searchLink> <br /><searchLink fieldCode="MM" term="%22Receptors%2C+CXCR%22">Receptors, CXCR*</searchLink>/<searchLink fieldCode="MM" term="%22Receptors%2C+CXCR+immunology%22">immunology</searchLink> <br /><searchLink fieldCode="MM" term="%22Receptors%2C+CXCR%22">Receptors, CXCR*</searchLink>/<searchLink fieldCode="MM" term="%22Receptors%2C+CXCR+genetics%22">genetics</searchLink><br /><searchLink fieldCode="MH" term="%22Animals%22">Animals</searchLink> ; <searchLink fieldCode="MH" term="%22Mice%22">Mice</searchLink> ; <searchLink fieldCode="MH" term="%22Immunization%22">Immunization</searchLink> ; <searchLink fieldCode="MH" term="%22Flow+Cytometry%22">Flow Cytometry</searchLink> ; <searchLink fieldCode="MH" term="%22Humans%22">Humans</searchLink>
– Name: Abstract
  Label: Abstract
  Group: Ab
  Data: Leukocyte migration is a fundamental process in both innate and adaptive immune responses. This process is tightly regulated by chemokines and their cognate receptors. The bioavailability of chemokines is further modulated by atypical chemokine receptors (ACKRs), a subset of chemokine receptor-like molecules that lack coupling to canonical G protein-mediated signaling pathways. Among these, ACKR4 regulates dendritic cell migration through ligand scavenging and has been implicated in tumor progression in murine models. We previously established anti-mouse ACKR4 (mACKR4) mAbs, A<subscript>4</subscript>Mab-1, A<subscript>4</subscript>Mab-2, and A<subscript>4</subscript>Mab-3, by N-terminal peptide immunization. This study examined the binding epitopes of A<subscript>4</subscript>Mabs. Alanine (or glycine) scanning within the N-terminal region (amino acids 2-19) was performed using flow cytometry and Western blotting. Results showed that Tyr12 is required for recognition by A<subscript>4</subscript>Mab-1 in flow cytometry, whereas Tyr11, Tyr12, Glu14, Glu15, and Glu17 are required in Western blotting. For A<subscript>4</subscript>Mab-2, Tyr12, Glu15, and Asn16 are required in flow cytometry, whereas Tyr11, Tyr12, Tyr13, Glu15, and Asn16 are required in Western blotting. Additionally, Glu14, Asn16, and Glu17 are required for recognition by A<subscript>4</subscript>Mab-3 in flow cytometry. These findings contribute to the understanding of mACKR4 recognition by A<subscript>4</subscript>Mabs.
– Name: Abstract
  Label: Competing Interests
  Group: Ab
  Data: Author Disclosure StatementThe authors have no conflicts of interest.
– Name: SubjectMinor
  Label: Contributed Indexing
  Group:
  Data: <i>Keywords: </i>alanine scanning; epitope mapping; flow cytometry; monoclonal antibody; mouse ACKR4
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  Label: Substance Nomenclature
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  Data: 0 (Antibodies, Monoclonal)<br />0 (Epitopes)<br />0 (Peptides)<br />0 (Receptors, CXCR)
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  Data: <i>Date Created: </i>20260616 <i>Date Completed: </i>20260702 <i>Latest Revision: </i>20260702
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  Data: 10.1177/21679436261461702
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  Data: 42300976
PLink https://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=cmedm&AN=42300976
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      – Type: doi
        Value: 10.1177/21679436261461702
    Languages:
      – Code: eng
        Text: English
    PhysicalDescription:
      Pagination:
        StartPage: 69
    Subjects:
      – SubjectFull: Animals
        Type: general
      – SubjectFull: Mice
        Type: general
      – SubjectFull: Immunization
        Type: general
      – SubjectFull: Flow Cytometry
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      – SubjectFull: Humans
        Type: general
      – SubjectFull: Epitope Mapping methods
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      – SubjectFull: Antibodies, Monoclonal immunology
        Type: general
      – SubjectFull: Epitopes immunology
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      – SubjectFull: Peptides immunology
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      – SubjectFull: Receptors, CXCR immunology
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      – SubjectFull: Receptors, CXCR genetics
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      – TitleFull: Epitope Mapping of Anti-mouse ACKR4 Monoclonal Antibodies Developed by N-Terminal Peptide Immunization.
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              Text: 2026 Jul-Aug
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              Y: 2026
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