Discover the SciOpen Platform and Achieve Your Research Goals with Ease.
Molecular imaging plays important roles in many fields, including disease diagnosis, therapeutic efficacy evaluation, intraoperative imaging guidance, drug metabolism monitoring, and patient selection for appropriate treatment. As a key component, the targeting ligand determines the specificity, affinity, and in vivo performance of molecular imaging probes. In this review, high‐throughput screening and biological display platforms for the discovery of ligands applicable to molecular imaging are briefly reviewed. Basic information on ligand development for molecular imaging is first introduced, followed by a presentation of various selection platforms and typical or iterative cases. The features, advantages, limitations, and application scope of screening and display platforms are compared and discussed. Last, a basic selection strategy and a perspective for protein‐based ligands are provided.
James ML, Gambhir SS. A molecular imaging primer: modalities, imaging agents, and applications. Physiol Rev. 2012;92(2):897–965. https://doi.org/10.1152/physrev.00049.2010
Rowe SP, Pomper MG. Molecular imaging in oncology: current impact and future directions. CA Cancer J Clin. 2022;72(4):333–52. https://doi.org/10.3322/caac.21713
Wu M, Shu J. Multimodal molecular imaging: current status and future directions. Contrast Media Mol Imaging. 2018;2018:1382183. https://doi.org/10.1155/2018/1382183
Hussain S, Mubeen I, Ullah N, Shah S, Khan BA, Zahoor M, et al. Modern diagnostic imaging technique applications and risk factors in the medical field: a review. BioMed Res Int. 2022;2022:5164970. https://doi.org/10.1155/2022/5164970
Chopra A, Shan L, Eckelman WC, Leung K, Latterner M, Bryant SH, et al. Molecular Imaging and Contrast Agent Database (MICAD): evolution and progress. Mol Imag Biol. 2012;14(1):4–13. https://doi.org/10.1007/s11307-011-0521-3
Lei Z, Ding L, Yao C, Mo F, Li C, Huang Y, et al. A highly efficient tumor‐targeting nanoprobe with a novel cell membrane permeability mechanism. Adv Mat. 2019;31(12):e1807456. https://doi.org/10.1002/adma.201807456
Charron CL, Hickey JL, Nsiama TK, Cruickshank DR, Turnbull WL, Luyt LG. Molecular imaging probes derived from natural peptides. Nat Prod Rep. 2016;33(6):761–800. https://doi.org/10.1039/c5np00083a
Ko YJ, Kim WJ, Kim K, Kwon IC. Advances in the strategies for designing receptor‐targeted molecular imaging probes for cancer research. J Contr Release. 2019;305:1–17. https://doi.org/10.1016/j.jconrel.2019.04.030
Zhao N, Qin Y, Liu H, Cheng Z. Tumor‐targeting peptides: ligands for molecular imaging and therapy. Anti Cancer Agents Med Chem. 2018;18(1):74–86. https://doi.org/10.2174/1871520617666170419143459
Cao R, Liu H, Cheng Z. Radiolabeled peptide probes for liver cancer imaging. Curr Med Chem. 2020;27(41):6968–86. https://doi.org/10.2174/0929867327666200320153837
Li X, Craven TW, Levine PM. Cyclic peptide screening methods for preclinical drug discovery. J Med Chem. 2022;65(18):11913–26. https://doi.org/10.1021/acs.jmedchem.2c01077
Luo R, Liu H, Cheng Z. Protein scaffolds: antibody alternatives for cancer diagnosis and therapy. RSC Chem Biol. 2022;3(7):830–47. https://doi.org/10.1039/d2cb00094f
Yang EY, Shah K. Nanobodies: next generation of cancer diagnostics and therapeutics. Front Oncol. 2020;10:1182. https://doi.org/10.3389/fonc.2020.01182
Jin S, Sun Y, Liang X, Gu X, Ning J, Xu Y, et al. Emerging new therapeutic antibody derivatives for cancer treatment. Signal Transduct Targeted Ther. 2022;7(1):39. https://doi.org/10.1038/s41392-021-00868-x
Liu R, Li X, Xiao W, Lam KS. Tumor‐targeting peptides from combinatorial libraries. Adv Drug Deliv Rev. 2017;110–111:13–37. https://doi.org/10.1016/j.addr.2016.05.009
Mahdavi SZB, Oroojalian F, Eyvazi S, Hejazi M, Baradaran B, Pouladi N, et al. An overview on display systems (phage, bacterial, and yeast display) for production of anticancer antibodies; advantages and disadvantages. Int J Biol Macromol. 2022;208:421–42. https://doi.org/10.1016/j.ijbiomac.2022.03.113
Zeng W, Guo L, Xu S, Chen J, Zhou J. High‐throughput screening technology in industrial biotechnology. Trends Biotechnol. 2020;38(8):888–906. https://doi.org/10.1016/j.tibtech.2020.01.001
Jaroszewicz W, Morcinek‐Orłowska J, Pierzynowska K, Gaffke L, Węgrzyn G. Phage display and other peptide display technologies. FEMS Microbiol Rev. 2022;46(2). https://doi.org/10.1093/femsre/fuab052
Saw PE, Song EW. Phage display screening of therapeutic peptide for cancer targeting and therapy. Protein Cell. 2019;10(11):787–807. https://doi.org/10.1007/s13238-019-0639-7
Sidhu SS. Engineering M13 for phage display. Biomol Eng. 2001;18(2):57–63. https://doi.org/10.1016/s1389-0344(01)00087-9
Deng X, Wang L, You X, Dai P, Zeng Y. Advances in the T7 phage display system (Review). Mol Med Rep. 2018;17(1):714–20. https://doi.org/10.3892/mmr.2017.7994
Brown KC. Peptidic tumor targeting agents: the road from phage display peptide selections to clinical applications. Curr Pharmaceut Des. 2010;16(9):1040–54. https://doi.org/10.2174/138161210790963788
Karasseva NG, Glinsky VV, Chen NX, Komatireddy R, Quinn TP. Identification and characterization of peptides that bind human ErbB‐2 selected from a bacteriophage display library. J Protein Chem. 2002;21(4):287–96. https://doi.org/10.1023/a:1019749504418
Shadidi M, Sioud M. Identification of novel carrier peptides for the specific delivery of therapeutics into cancer cells. Faseb J. 2003;17(2):256–8. https://doi.org/10.1096/fj.02-0280fje
Ringhieri P, Mannucci S, Conti G, Nicolato E, Fracasso G, Marzola P, et al. Liposomes derivatized with multimeric copies of KCCYSL peptide as targeting agents for HER‐2‐overexpressing tumor cells. Int J Nanomed. 2017;12:501–14. https://doi.org/10.2147/ijn.s113607
Xing L, Xu Y, Sun K, Wang H, Zhang F, Zhou Z, et al. Identification of a peptide for folate receptor alpha by phage display and its tumor targeting activity in ovary cancer xenograft. Sci Rep. 2018;8(1):8426. https://doi.org/10.1038/s41598-018-26683-z
Pola R, Böhmová E, Filipová M, Pechar M, Pankrác J, Větvička D, et al. Targeted polymer‐based probes for fluorescence guided visualization and potential surgery of EGFR‐positive head‐and‐neck tumors. Pharmaceutics. 2020;12(1):31. https://doi.org/10.3390/pharmaceutics12010031
Joshi BP, Dai Z, Gao Z, Lee JH, Ghimire N, Chen J, et al. Detection of sessile serrated adenomas in the proximal colon using wide‐field fluorescence endoscopy. Gastroenterology. 2017;152(5):1002–13.e9. https://doi.org/10.1053/j.gastro.2016.12.009
Zhang D, Huang J, Li W, Zhang Z, Zhu M, Feng Y, et al. Screening and identification of a CD44v6 specific peptide using improved phage display for gastric cancer targeting. Ann Transl Med. 2020;8(21):1442. https://doi.org/10.21037/atm-19-4781
Zhang H, Chen S. Cyclic peptide drugs approved in the last two decades (2001–2021). RSC Chem Biol. 2022;3(1):18–31. https://doi.org/10.1039/d1cb00154j
Shinbara K, Liu W, van Neer RHP, Katoh T, Suga H. Methodologies for backbone macrocyclic peptide synthesis compatible with screening technologies. Front Chem. 2020;8:447. https://doi.org/10.3389/fchem.2020.00447
Owens AE, Iannuzzelli JA, Gu Y, Fasan R. MOrPH‐PhD: an integrated phage display platform for the discovery of functional genetically encoded peptide macrocycles. ACS Cent Sci. 2020;6(3):368–81. https://doi.org/10.1021/acscentsci.9b00927
Chen S, Lovell S, Lee S, Fellner M, Mace PD, Bogyo M. Identification of highly selective covalent inhibitors by phage display. Nat Biotechnol. 2021;39(4):490–8. https://doi.org/10.1038/s41587-020-0733-7
Cho HJ, Park SJ, Lee YS, Kim S. Theranostic iRGD peptide containing cisplatin prodrug: dual‐cargo tumor penetration for improved imaging and therapy. J Contr Release. 2019;300:73–80. https://doi.org/10.1016/j.jconrel.2019.02.043
Burggraaf J, Kamerling IM, Gordon PB, Schrier L, de Kam ML, Kales AJ, et al. Detection of colorectal polyps in humans using an intravenously administered fluorescent peptide targeted against c‐Met. Nat Med. 2015;21(8):955–61. https://doi.org/10.1038/nm.3641
Ye XX, Zhao YY, Wang Q, Xiao W, Zhao J, Peng YJ, et al. EDB fibronectin‐specific SPECT probe (99m)Tc‐HYNIC‐ZD2 for breast cancer detection. ACS Omega. 2017;2(6):2459–68. https://doi.org/10.1021/acsomega.7b00226
Yan J, Yu X, Chen X, Liu F, Chen F, Ding N, et al. Identification of a glypican‐3 binding peptide from a phage‐displayed peptide library for PET imaging of hepatocellular carcinoma. Front Oncol. 2021;11:679336. https://doi.org/10.3389/fonc.2021.679336
Yang X, Li Y, Zhu Z, Huang X, Wang T, Yuan J, et al. Identification of a peptide that crosses the blood‐cerebrospinal fluid barrier by phage display technology. Amino Acids. 2021;53(8):1181–6. https://doi.org/10.1007/s00726-021-03016-5
Ståhl S, Gräslund T, Eriksson Karlström A, Frejd FY, Nygren P, Löfblom J. Affibody molecules in biotechnological and medical applications. Trends Biotechnol. 2017;35(8):691–712. https://doi.org/10.1016/j.tibtech.2017.04.007
Wikman M, Steffen AC, Gunneriusson E, Tolmachev V, Adams GP, Carlsson J, et al. Selection and characterization of HER2/neu‐binding affibody ligands. Protein Eng Des Sel. 2004;17(5):455–62. https://doi.org/10.1093/protein/gzh053
Orlova A, Magnusson M, Eriksson TL, Nilsson M, Larsson B, Höidén‐Guthenberg I, et al. Tumor imaging using a picomolar affinity HER2 binding affibody molecule. Cancer Res. 2006;66(8):4339–48. https://doi.org/10.1158/0008-5472.can-05-3521
Baum RP, Prasad V, Müller D, Schuchardt C, Orlova A, Wennborg A, et al. Molecular imaging of HER2‐expressing malignant tumors in breast cancer patients using synthetic 111In‐ or 68Ga‐labeled affibody molecules. J Nucl Med. 2010;51(6):892–7. https://doi.org/10.2967/jnumed.109.073239
Feldwisch J, Tolmachev V, Lendel C, Herne N, Sjöberg A, Larsson B, et al. Design of an optimized scaffold for affibody molecules. J Mol Biol. 2010;398(2):232–47. https://doi.org/10.1016/j.jmb.2010.03.002
Sörensen J, Velikyan I, Sandberg D, Wennborg A, Feldwisch J, Tolmachev V, et al. Measuring HER2‐receptor expression in metastatic breast cancer using [68Ga]ABY‐025 affibody PET/CT. Theranostics. 2016;6(2):262–71. https://doi.org/10.7150/thno.13502
Woldring DR, Holec PV, Stern LA, Du Y, Hackel BJ. A gradient of sitewise diversity promotes evolutionary fitness for binder discovery in a three‐helix bundle protein scaffold. Biochemistry. 2017;56(11):1656–71. https://doi.org/10.1021/acs.biochem.6b01142
Moreno E, Valdés‐Tresanco MS, Molina‐Zapata A, Sánchez‐Ramos O. Structure‐based design and construction of a synthetic phage display nanobody library. BMC Res Notes. 2022;15(1):124. https://doi.org/10.1186/s13104-022-06001-7
Liu B, Yang D. Easily established and multifunctional synthetic nanobody libraries as research tools. Int J Mol Sci. 2022;23(3):1482. https://doi.org/10.3390/ijms23031482
Ma J, Xu X, Fu C, Xia P, Tian M, Zheng L, et al. CDH17 nanobodies facilitate rapid imaging of gastric cancer and efficient delivery of immunotoxin. Biomater Res. 2022;26(1):64. https://doi.org/10.1186/s40824-022-00312-3
Yu X, Long Y, Chen B, Tong Y, Shan M, Jia X, et al. PD‐L1/TLR7 dual‐targeting nanobody‐drug conjugate mediates potent tumor regression via elevating tumor immunogenicity in a host‐expressed PD‐L1 bias‐dependent way. J Immunother Cancer. 2022;10(10):e004590. https://doi.org/10.1136/jitc-2022-004590
Maruthachalam BV, El‐Sayed A, Liu J, Sutherland AR, Hill W, Alam MK, et al. A single‐framework synthetic antibody library containing a combination of canonical and variable complementarity‐determining regions. Chembiochem. 2017;18(22):2247–59. https://doi.org/10.1002/cbic.201700279
El‐Sayed A, Bernhard W, Barreto K, Gonzalez C, Hill W, Pastushok L, et al. Evaluation of antibody fragment properties for near‐infrared fluorescence imaging of HER3‐positive cancer xenografts. Theranostics. 2018;8(17):4856–69. https://doi.org/10.7150/thno.24252
Hintz HM, Cowan AE, Shapovalova M, LeBeau AM. Development of a cross‐reactive monoclonal antibody for detecting the tumor stroma. Bioconjugate Chem. 2019;30(5):1466–76. https://doi.org/10.1021/acs.bioconjchem.9b00206
Diebolder P, Mpoy C, Scott J, Huynh TT, Fields R, Spitzer D, et al. Preclinical evaluation of an engineered single‐chain fragment variable‐fragment crystallizable targeting human CD44. J Nucl Med. 2021;62(1):137–43. https://doi.org/10.2967/jnumed.120.249557
Löfblom J. Bacterial display in combinatorial protein engineering. Biotechnol J. 2011;6(9):1115–29. https://doi.org/10.1002/biot.201100129
Mazor Y, Van Blarcom T, Iverson BL, Georgiou G. Isolation of full‐length IgG antibodies from combinatorial libraries expressed in Escherichia coli. Methods Mol Biol. 2009;525:217–39, xiv. https://doi.org/10.1007/978-1-59745-554-1_11
Friedrich L, Kornberger P, Mendler CT, Multhoff G, Schwaiger M, Skerra A. Selection of an Anticalin® against the membrane form of Hsp70 via bacterial surface display and its theranostic application in tumour models. Biol Chem. 2018;399(3):235–52. https://doi.org/10.1515/hsz-2017-0207
Kronqvist N, Malm M, Göstring L, Gunneriusson E, Nilsson M, Höidén Guthenberg I, et al. Combining phage and staphylococcal surface display for generation of ErbB3‐specific affibody molecules. Protein Eng Des Sel. 2011;24(4):385–96. https://doi.org/10.1093/protein/gzq118
Malm M, Kronqvist N, Lindberg H, Gudmundsdotter L, Bass T, Frejd FY, et al. Inhibiting HER3‐mediated tumor cell growth with affibody molecules engineered to low picomolar affinity by position‐directed error‐prone PCR‐like diversification. PLoS One. 2013;8(5):e62791. https://doi.org/10.1371/journal.pone.0062791
Dahlsson Leitao C, Rinne SS, Mitran B, Vorobyeva A, Andersson KG, Tolmachev V, et al. Molecular design of HER3‐targeting affibody molecules: influence of chelator and presence of HEHEHE‐tag on biodistribution of (68)Ga‐labeled tracers. Int J Mol Sci. 2019;20(5):1080. https://doi.org/10.3390/ijms20051080
Fleetwood F, Klint S, Hanze M, Gunneriusson E, Frejd FY, Ståhl S, et al. Simultaneous targeting of two ligand‐binding sites on VEGFR2 using biparatopic affibody molecules results in dramatically improved affinity. Sci Rep. 2014;4(1):7518. https://doi.org/10.1038/srep07518
Fleetwood F, Güler R, Gordon E, Ståhl S, Claesson‐Welsh L, Löfblom J. Novel affinity binders for neutralization of vascular endothelial growth factor (VEGF) signaling. Cell Mol Life Sci. 2016;73(8):1671–83. https://doi.org/10.1007/s00018-015-2088-7
Mitran B, Güler R, Roche FP, Lindström E, Selvaraju RK, Fleetwood F, et al. Radionuclide imaging of VEGFR2 in glioma vasculature using biparatopic affibody conjugate: proof‐of‐principle in a murine model. Theranostics. 2018;8(16):4462–76. https://doi.org/10.7150/thno.24395
Teymennet‐Ramírez KV, Martínez‐Morales F, Trejo‐Hernández MR. Yeast surface display system: strategies for improvement and biotechnological applications. Front Bioeng Biotechnol. 2021;9:794742. https://doi.org/10.3389/fbioe.2021.794742
Könning D, Kolmar H. Beyond antibody engineering: directed evolution of alternative binding scaffolds and enzymes using yeast surface display. Microb Cell Factories. 2018;17(1):32. https://doi.org/10.1186/s12934-018-0881-3
Bam R, Lown PS, Stern LA, Sharma K, Wilson KE, Bean GR, et al. Efficacy of affibody‐based ultrasound molecular imaging of vascular B7‐H3 for breast cancer detection. Clin Cancer Res. 2020;26(9):2140–50. https://doi.org/10.1158/1078-0432.ccr-19-1655
Case BA, Kruziki MA, Johnson SM, Hackel BJ. Engineered charge redistribution of Gp2 proteins through guided diversity for improved PET imaging of epidermal growth factor receptor. Bioconjugate Chem. 2018;29(5):1646–58. https://doi.org/10.1021/acs.bioconjchem.8b00144
Lu Z, Kamat K, Johnson BP, Yin CC, Scholler N, Abbott KL. Generation of a fully human scFv that binds tumor‐specific glycoforms. Sci Rep. 2019;9(1):5101. https://doi.org/10.1038/s41598-019-41567-6
Abou‐Elkacem L, Wang H, Chowdhury SM, Kimura RH, Bachawal SV, Gambhir SS, et al. Thy1‐Targeted microbubbles for ultrasound molecular imaging of pancreatic ductal adenocarcinoma. Clin Cancer Res. 2018;24(7):1574–85. https://doi.org/10.1158/1078-0432.ccr-17-2057
Kimura RH, Jones DS, Jiang L, Miao Z, Cheng Z, Cochran JR. Functional mutation of multiple solvent‐exposed loops in the Ecballium elaterium trypsin inhibitor‐II cystine knot miniprotein. PLoS One. 2011;6(2):e16112. https://doi.org/10.1371/journal.pone.0016112
Jiang L, Zhu H, Li Y, Wu X, Wang H, Cheng Z. Detecting vulnerable atherosclerotic plaques by (68)Ga‐labeled divalent cystine knot peptide. Mol Pharm. 2019;16(3):1350–7. https://doi.org/10.1021/acs.molpharmaceut.8b01291
Kimura RH, Wang L, Shen B, Huo L, Tummers W, Filipp FV, et al. Evaluation of integrin αvβ(6) cystine knot PET tracers to detect cancer and idiopathic pulmonary fibrosis. Nat Commun. 2019;10(1):4673. https://doi.org/10.1038/s41467-019-11863-w
Mattheakis LC, Bhatt RR, Dower WJ. An in vitro polysome display system for identifying ligands from very large peptide libraries. Proc Natl Acad Sci USA. 1994;91(19):9022–6. https://doi.org/10.1073/pnas.91.19.9022
Plückthun A. Ribosome display: a perspective. Methods Mol Biol. 2012;805:3–28. https://doi.org/10.1007/978-1-61779-379-0_1
Lagoutte P. [Ribosome display: evolution and acellular selection of molecular libraries for high affinity binder generation]. M‐S (Med Sci). 2020;36(8–9):717–24. https://doi.org/10.1051/medsci/2020126
Li R, Kang G, Hu M, Huang H. Ribosome display: a potent display technology used for selecting and evolving specific binders with desired properties. Mol Biotechnol. 2019;61(1):60–71. https://doi.org/10.1007/s12033-018-0133-0
Petroková H, Mašek J, Kuchař M, Vítečková Wünschová A, Štikarová J, Bartheldyová E, et al. Targeting human thrombus by liposomes modified with anti‐fibrin protein binders. Pharmaceutics. 2019;11(12):642. https://doi.org/10.3390/pharmaceutics11120642
Kosareva A, Punjabi M, Ochoa‐Espinosa A, Xu L, Schaefer JV, Dreier B, et al. Designed Ankyrin repeat proteins as novel binders for ultrasound molecular imaging. Ultrasound Med Biol. 2021;47(9):2664–75. https://doi.org/10.1016/j.ultrasmedbio.2021.04.027
Kamalinia G, Grindel BJ, Takahashi TT, Millward SW, Roberts RW. Directing evolution of novel ligands by mRNA display. Chem Soc Rev. 2021;50(16):9055–103. https://doi.org/10.1039/d1cs00160d
Li S, Millward S, Roberts R. In vitro selection of mRNA display libraries containing an unnatural amino acid. J Am Chem Soc. 2002;124(34):9972–3. https://doi.org/10.1021/ja026789q
Alleyne C, Amin RP, Bhatt B, Bianchi E, Blain JC, Boyer N, et al. Series of novel and highly potent cyclic peptide PCSK9 inhibitors derived from an mRNA display screen and optimized via structure‐based design. J Med Chem. 2020;63(22):13796–824. https://doi.org/10.1021/acs.jmedchem.0c01084
Grindel BJ, Engel BJ, Ong JN, Srinivasamani A, Liang X, Zacharias NM, et al. Directed evolution of PD‐L1‐targeted affibodies by mRNA display. ACS Chem Biol. 2022;17(6):1543–55. https://doi.org/10.1021/acschembio.2c00218
Fiacco SV, Kelderhouse LE, Hardy A, Peleg Y, Hu B, Ornelas A, et al. Directed evolution of scanning unnatural‐protease‐resistant (SUPR) peptides for in vivo applications. Chembiochem. 2016;17(17):1643–51. https://doi.org/10.1002/cbic.201600253
Kamalinia G, Engel BJ, Srinivasamani A, Grindel BJ, Ong JN, Curran MA, et al. mRNA display discovery of a novel programmed death ligand 1 (PD‐L1) binding peptide (a peptide ligand for PD‐L1). ACS Chem Biol. 2020;15(6):1630–41. https://doi.org/10.1021/acschembio.0c00264
Sakai K, Passioura T, Sato H, Ito K, Furuhashi H, Umitsu M, et al. Macrocyclic peptide‐based inhibition and imaging of hepatocyte growth factor. Nat Chem Biol. 2019;15(6):598–606. https://doi.org/10.1038/s41589-019-0285-7
This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.