PMID- 37553068 OWN - NLM STAT- MEDLINE DCOM- 20230821 LR - 20240210 IS - 2161-5063 (Electronic) IS - 2161-5063 (Linking) VI - 12 IP - 8 DP - 2023 Aug 18 TI - A Cell-Free Protein Synthesis Platform to Produce a Clinically Relevant Allergen Panel. PG - 2252-2261 LID - 10.1021/acssynbio.3c00269 [doi] AB - Allergens are used in the clinical diagnosis (e.g., skin tests) and treatment (e.g., immunotherapy) of allergic diseases. With growing interest in molecular allergy diagnostics and precision therapies, new tools are needed for producing allergen-based reagents. As a step to address this need, we demonstrate a cell-free protein synthesis approach for allergen production of a clinically relevant allergen panel composed of common allergens spanning a wide range of phylogenetic kingdoms. We show that allergens produced with this approach can be recognized by allergen-specific immunoglobulin E (IgE), either monoclonals or in patient sera. We also show that a cell-free expressed allergen can activate human cells such as peripheral blood basophils and CD34+ progenitor-derived mast cells in an IgE-dependent manner. We anticipate that this cell-free platform for allergen production will enable diagnostic and therapeutic technologies, providing useful tools and treatments for both the allergist and allergic patient. FAU - Thames, Ariel Helms AU - Thames AH AUID- ORCID: 0000-0001-5418-6048 AD - Medical Scientist Training Program, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, United States. AD - Center for Synthetic Biology, Northwestern University, 2145 Sheridan Road, Tech B486, Evanston, Illinois 60208, United States. AD - Interdisciplinary Biological Sciences Program, Northwestern University, Evanston, Illinois 60208, United States. AD - Division of Allergy and Immunology, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, United States. FAU - Rische, Clayton H AU - Rische CH AUID- ORCID: 0000-0003-3610-2116 AD - Center for Synthetic Biology, Northwestern University, 2145 Sheridan Road, Tech B486, Evanston, Illinois 60208, United States. AD - Division of Allergy and Immunology, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, United States. AD - Department of Chemical and Biological Engineering, Northwestern University, 2145 Sheridan Rd, Evanston, Illinois 60208, United States. AD - Simpson Querrey Institute, Northwestern University, Chicago, Illinois 60611, United States. FAU - Cao, Yun AU - Cao Y AD - Division of Allergy and Immunology, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, United States. FAU - Krier-Burris, Rebecca A AU - Krier-Burris RA AD - Division of Allergy and Immunology, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, United States. FAU - Kuang, Fei Li AU - Kuang FL AD - Division of Allergy and Immunology, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, United States. FAU - Hamilton, Robert G AU - Hamilton RG AD - Division of Allergy and Clinical Immunology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland 21224, United States. FAU - Bronzert, Charles AU - Bronzert C AD - Division of Allergy and Clinical Immunology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland 21224, United States. FAU - Bochner, Bruce S AU - Bochner BS AUID- ORCID: 0000-0002-9411-6009 AD - Medical Scientist Training Program, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, United States. AD - Division of Allergy and Immunology, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, United States. FAU - Jewett, Michael C AU - Jewett MC AUID- ORCID: 0000-0003-2948-6211 AD - Medical Scientist Training Program, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, United States. AD - Center for Synthetic Biology, Northwestern University, 2145 Sheridan Road, Tech B486, Evanston, Illinois 60208, United States. AD - Interdisciplinary Biological Sciences Program, Northwestern University, Evanston, Illinois 60208, United States. AD - Department of Chemical and Biological Engineering, Northwestern University, 2145 Sheridan Rd, Evanston, Illinois 60208, United States. AD - Simpson Querrey Institute, Northwestern University, Chicago, Illinois 60611, United States. AD - Department of Bioengineering, Stanford University, Stanford, California 94305, United States. LA - eng GR - F31 AI165279/AI/NIAID NIH HHS/United States GR - K23 AI171085/AI/NIAID NIH HHS/United States PT - Letter PT - Research Support, N.I.H., Extramural PT - Research Support, U.S. Gov't, Non-P.H.S. DEP - 20230808 PL - United States TA - ACS Synth Biol JT - ACS synthetic biology JID - 101575075 RN - 0 (Allergens) RN - 37341-29-0 (Immunoglobulin E) SB - IM MH - Humans MH - *Allergens MH - Phylogeny MH - *Immunoglobulin E PMC - PMC10768853 MID - NIHMS1952288 OTO - NOTNLM OT - IgE recognition OT - allergen OT - basophils OT - cell-free protein synthesis OT - mast cells COIS- The authors declare the following competing financial interest(s): B.S.B. receives remuneration for serving on the scientific advisory board of Allakos, Inc. and owns stock in Allakos. He receives consulting fees from Third Harmonic Bio, Lupagen, Sanofi, and Acelyrin. He receives publication-related royalty payments from Elsevier and UpToDate. He is a co-inventor on existing Siglec-8-related patents and thus may be entitled to a share of royalties received by Johns Hopkins University during development and potential sales of such products. B.S.B. is also a co-founder of Allakos, Inc. which makes him subject to certain restrictions under university policy. The terms of this arrangement are being managed by Johns Hopkins University and Northwestern University in accordance with their conflict-of-interest policies. M.C.J. has a financial interest in SwiftScale Biologics, Gauntlet Bio, Pearl Bio, Inc., Design Pharmaceutics, and Stemloop Inc. M.C.J.s interests are reviewed and managed by Northwestern University and Stanford University in accordance with their competing interest policies. All other authors declare no competing interests. The authors have filed an invention disclosure based on the work presented. EDAT- 2023/08/09 01:05 MHDA- 2023/08/21 06:42 PMCR- 2024/08/18 CRDT- 2023/08/08 19:12 PHST- 2024/08/18 00:00 [pmc-release] PHST- 2023/08/21 06:42 [medline] PHST- 2023/08/09 01:05 [pubmed] PHST- 2023/08/08 19:12 [entrez] AID - 10.1021/acssynbio.3c00269 [doi] PST - ppublish SO - ACS Synth Biol. 2023 Aug 18;12(8):2252-2261. doi: 10.1021/acssynbio.3c00269. Epub 2023 Aug 8.