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JP-7857228-B2 - Antibodies that specifically bind to peptides associated with primary immunodeficiency disorders, namely Wiscott-Aldrich syndrome and X-linked agammaglobulinemia.

JP7857228B2JP 7857228 B2JP7857228 B2JP 7857228B2JP-7857228-B2

Inventors

  • ハーン，シフーン
  • コリンズ，クリストファー
  • イー，ファン
  • ダユハ，レムウィリン

Assignees

  • シアトル・チルドレンズ・ホスピタル・ドゥーイング／ビジネス／アズ・シアトル・チルドレンズ・リサーチ・インスティテュート

Dates

Publication Date
20260512
Application Date
20210402
Priority Date
20200402

Claims (20)

  1. A heavy chain variable (VH) domain comprising CDRH1 having the sequence shown in SEQ ID NO: 3, CDRH2 having the sequence shown in SEQ ID NO: 4, and CDRH3 having the sequence shown in SEQ ID NO: 5, An antibody or antigen-binding fragment thereof that binds to a WASp 289 signature peptide, comprising a light chain variable (VL) domain including CDRL1 having the sequence shown in SEQ ID NO: 6, CDRL2 having the sequence shown in SEQ ID NO: 7, and CDRL3 having the sequence shown in SEQ ID NO: 8.
  2. The antibody or antigen-binding fragment according to claim 1, comprising one or more of the following: a VH domain having the sequence shown in SEQ ID NO: 15; a heavy chain having the sequence shown in SEQ ID NO: 17; a VL domain having the sequence shown in SEQ ID NO: 16; or a light chain having the sequence shown in SEQ ID NO: 18.
  3. The VH domain has the sequence shown in Sequence ID 15, and the VL domain has the sequence shown in Sequence ID 16, or The heavy chain has the sequence shown in Sequence ID 17, and the light chain has the sequence shown in Sequence ID 18. The antibody or antigen-binding fragment thereof according to claim 1 or 2.
  4. A process of digesting proteins derived from biological samples obtained from the subject with enzymes to produce one or more peptides, A step of concentrating the WASp signature peptide with the antibody or antigen-binding fragment thereof according to claim 1, A method for providing data for the diagnosis of primary immunodeficiency disorders (PIDDs), comprising the step of determining the concentration of a WASp signature peptide by performing liquid chromatography-multi-reaction monitoring mass spectrometry (LC-MRM-MS) on a concentrated WASp signature peptide.
  5. The process further includes comparing the concentration of the WASp signature peptide with a predetermined threshold concentration. The method according to claim 4, wherein if the concentration of the WASp signature peptide is lower than the predetermined threshold concentration, or if the WASp signature peptide is not present, the subject is suspected to have Wiscott-Aldrich syndrome (WAS).
  6. The method according to claim 5, wherein a predetermined threshold concentration of the WASp signature peptide is calculated from the standard deviation of the mean concentration of the WASp signature peptide in biological samples derived from a population of normal control subjects.
  7. The method according to claim 6, wherein the biological sample is DBS, and the average concentration of the WASp signature peptide in DBS derived from a population of normal control subjects includes concentrations in the range of 7,000 pmol/L to 30,000 pmol/L.
  8. A step of concentrating the BTK signature peptide with an antibody that binds to BTK545 or its antigen-binding fragment, The method according to any one of claims 4 to 7, further comprising the step of performing liquid chromatography-multi-reaction monitoring mass spectrometry (LC-MRM-MS) on the concentrated BTK peptide to determine the concentration of the BTK signature peptide.
  9. An antibody or its antigen-binding fragment that binds to BTK545, The method according to claim 8, comprising a VH domain including CDRH1 having the sequence shown in SEQ ID NO: 9, CDRH2 having the sequence shown in SEQ ID NO: 10, and CDRH3 having the sequence shown in SEQ ID NO: 11, and a VL domain including CDRL1 having the sequence shown in SEQ ID NO: 12, CDRL2 having the sequence shown in SEQ ID NO: 13, and CDRL3 having the sequence shown in SEQ ID NO: 14.
  10. An antibody or its antigen-binding fragment that binds to BTK545, (a) A VH domain having the sequence shown in Sequence ID No. 23, and a VL domain having the sequence shown in Sequence ID No. 24, or (b) The method according to claim 9, comprising a heavy chain having the sequence shown in Sequence ID No. 25 and a light chain having the sequence shown in Sequence ID No. 26.
  11. The biological sample is a dried blood spot (DBS), an oral swab, peripheral blood mononuclear cells (PBMCs), or white blood cells (WBCs), and/or The method according to any one of claims 4 to 10, wherein the enzyme is trypsin.
  12. The process further includes comparing the concentration of the BTK signature peptide with a predetermined threshold concentration. The method according to any one of claims 8 to 11, wherein if the concentration of the BTK signature peptide is lower than the predetermined threshold concentration, or if the BTK signature peptide is not present, the subject is suspected to be suffering from X-linked agammaglobulinemia (XLA).
  13. The method according to any one of claims 4 to 12, performed as part of a neonatal screening (NBS) to further screen the subject for one or more of the following conditions: phenylketonuria, primary congenital hypothyroidism, cystic fibrosis, and sickle cell anemia.
  14. The method according to any one of claims 4 to 13, performed in a state in which the subject is free from clinical symptoms of WAS and/or XLA.
  15. The method according to claim 12, wherein the predetermined threshold concentration of the BTK signature peptide is calculated from the standard deviation of the mean concentration of the BTK signature peptide in biological samples derived from a population of normal control subjects.
  16. The method according to claim 15, wherein the average concentration of the BTK signature in DBS derived from a normal control population includes concentrations in the range of 400 pmol/L to 2000 pmol/L.
  17. A kit for assays for screening Wiscott-Aldrich syndrome (WAS) in subjects, (i) an antibody or antigen-binding fragment thereof as described in claim 1, and (ii) a reference signature peptide comprising the WASp signature peptide of the WAS of SEQ ID NO: 1, for assay.
  18. The assay kit according to claim 17, further comprising (iii) an antibody or antigen-binding fragment thereof that binds to BTK545, and (iv) a reference signature peptide comprising the BTK signature peptide of XLA of SEQ ID NO: 2, for the screening of Wiscott-Aldrich syndrome (WAS) and X-linked agammaglobulinemia (XLA) in a subject .
  19. The assay kit according to claim 17 or 18, wherein the reference signature peptide is isotope-labeled, and/or the antibody or its antigen-binding fragment is attached to a magnetic bead.
  20. The assay kit according to any one of claims 17 to 19, further comprising one or more additional components selected from a filter paper card, an oral cotton swab, a blood collection tube, a punch tool, a digestive enzyme, a digestive buffer, a solid support for an antibody or its antigen-binding fragment, and an elution buffer .

Description

Cross-reference to Related Applications This application claims priority to U.S. Provisional Patent Application No. 63/004,415, filed 2 April 2020, which is incorporated herein by reference in its entirety as if it were fully described herein. Federal Government Sponsored Research or Development Statement: This invention was made with government support under AI 123135, awarded by the National Institutes of Health. The government reserves certain rights in this invention. Statement Regarding Sequence Listing The sequence listing relating to this application is provided in text format instead of a paper copy and is incorporated herein by reference. The name of the text file containing the sequence listing is 2GV8205_ST25.txt. The text file is 59KB in size, was created on April 2, 2021, and filed electronically via EFS-Web. Area of Disclosure The current disclosure provides antibodies that bind to peptides associated with primary immunodeficiency disorders (PIDDs), such as Wiscott-Aldrich syndrome and X-linked agammaglobulinemia (XLA). Among other applications, these antibodies may be used in the clinical diagnosis and neonatal screening of WAS and XLA. Many diseases have effective treatments available. However, in many of these diseases, once symptoms appear, the disease is already fatal or causes irreversible damage. An example of such a disease is primary immunodeficiency disorder (PIDD). Primary immunodeficiency disorders (PIDDs), also known as congenital immunodeficiency disorders (IEIs), are a group of more than 416 rare genetic disorders in which components of the immune system are missing or improperly functioning. Examples of PIDDs include Wiscott-Aldrich syndrome (WAS) and X-linked agammaglobulinemia (XLA). Early detection of PIDDs is crucial for controlling and preventing potentially life-threatening infections and chronic sequelae. If a diagnosis could be made before clinical symptoms appeared, the management of PIDD would be significantly enhanced. Newborn screening (NBS) is a standard, public, preventative, and mandatory screening test for the 4 million infants born annually in the United States. NBS typically involves a blood test performed 24 to 48 hours after birth. The screening uses a few drops of blood from the newborn's heel, collected on filter paper. The paper containing the dried blood spot (DBS) can be stored until the test is performed. To perform an NBS assessment, a punch of dried blood is taken from the DBS (Database for Sickle Cell Biology) and clinical tests are performed to detect the presence of specific substances in the blood (called markers or biomarkers) that indicate disorders that are not apparent at birth but can cause serious health problems later in life. The disorders screened vary by state, but most states screen for phenylketonuria, primary congenital hypothyroidism, cystic fibrosis, and sickle cell anemia. NBS has proven highly effective in improving patient outcomes, preventing long-term disability in affected individuals, and reducing healthcare costs. International Patent Application No. PCT/US2019/054856 describes the development of a multiplexed assay that may be used to screen newborns for severe combined immunodeficiency (SCID), Wiscott-Aldrich syndrome (WAS), X-linked agammaglobulinemia (XLA), cystinosis, and Wilson's disease (WD). This assay may significantly improve outcomes for affected individuals by reliably diagnosing these disorders before the onset of clinically significant and often fatal symptoms. The assay may detect the presence or absence of markers associated with these disorders using dried blood spots (DBS), which are already conventionally collected as part of existing neonatal screening (NBS) procedures. The multiplexed assay described in PCT/US2019/054856 utilizes peptide immunoaffinity enrichment combined with selective reaction monitoring mass spectrometry (immuno-SRM). This disclosure provides antibodies that specifically bind to signature peptides associated with primary immunodeficiency disorders (PIDDs), namely Wiscott-Aldrich syndrome (WAS) and X-linked agammaglobulinemia (XLA). Specific embodiments include an antibody or antigen-binding fragment that binds to the WASp 289 signature peptide of WAS (SEQ ID NO: 1), and comprises: a heavy chain variable (VH) domain including CDRH1 of SEQ ID NO: 3, CDRH2 of SEQ ID NO: 4, and CDRH3 of SEQ ID NO: 5, and a light chain variable (VL) domain including CDRL1 of SEQ ID NO: 6, CDRL2 of SEQ ID NO: 7, and CDRL3 of SEQ ID NO: 8. In specific embodiments, the antibody or antigen-binding fragment that binds to the WASp 289 signature peptide includes a VH domain including SEQ ID NO: 15. In specific embodiments, the antibody or antigen-binding fragment that binds to the WASp 289 signature peptide includes a VL domain including SEQ ID NO: 16. In specific embodiments, the antibody or antigen-binding fragment that binds to the WASp 289 signature peptide includes a heavy chain including SEQ ID NO: 17. In