Abstract
Immunocapture is now a well-established method for sample preparation prior to quantitation of peptides and proteins in complex matrices. This short review will give an overview of some clinical applications of immunocapture methods, as well as protocols with and without enzymatic digestion in a clinical context. The advantages and limitations of both approaches are discussed in detail. Challenges related to the choice of mass spectrometer are also discussed. Top-down, middle-down, and bottom-up approaches are discussed. Even though immunocapture has its limitations, its main advantage is that it provides an additional dimension of separation and/or isolation when working with peptides and proteins. Overall, this short review demonstrates the potential of such techniques in the field of proteomics-based clinical medicine and paves the way for better personalized medicine.
-
Research funding: None declared.
-
Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.
-
Competing interests: Authors state no conflict of interest.
-
Informed consent: Not applicable.
-
Ethical approval: Not applicable.
References
1. Rosenfeld, L. Four centuries of clinical chemistry. London, UK: Routledge; 2018.10.1201/9780203755525Search in Google Scholar
2. Antonelli, G, Ceccato, F, Artusi, C, Marinova, M, Plebani, M. Salivary cortisol and cortisone by LC–MS/MS: validation, reference intervals and diagnostic accuracy in cushing’s syndrome. Clin Chim Acta 2015;451:247–51. https://doi.org/10.1016/j.cca.2015.10.004.Search in Google Scholar PubMed
3. Jones, G, Kaufmann, M. Vitamin D metabolite profiling using liquid chromatography–tandem mass spectrometry (LC–MS/MS). J Steroid Biochem Mol Biol 2016;164:110–4. https://doi.org/10.1016/j.jsbmb.2015.09.026.Search in Google Scholar PubMed
4. Yu, S, Wang, D, Ma, X, Zhang, Y, Sun, D, Zou, Y, et al.. Analytical and clinical performance of a liquid chromatography–tandem mass spectrometry method for measuring gastrin subtypes G34 and G17 in serum. Clin Chem 2021;67:1220–9. https://doi.org/10.1093/clinchem/hvab097.Search in Google Scholar PubMed
5. Grebe, SKG, Singh, RJ. LC-MS/MS in the clinical laboratory – where to from here? Clin Biochem Rev 2011;32:5–31.Search in Google Scholar
6. Le Goff, C, Farre-Segura, J, Stojkovic, V, Dufour, P, Peeters, S, Courtois, J, et al.. The pathway through LC-MS method development: in-house or ready-to-use kit-based methods? Clin Chem Lab Med 2020;58:1002–9. https://doi.org/10.1515/cclm–2019–0916.10.1515/cclm-2019-0916Search in Google Scholar PubMed
7. Kellie, JF, Tran, JC, Jian, W, Jones, B, Mehl, JT, Ge, Y, et al.. Intact protein mass spectrometry for therapeutic protein quantitation, pharmacokinetics, and biotransformation in preclinical and clinical studies: an industry perspective. J Am Soc Mass Spectrom 2021;32:1886–900. https://doi.org/10.1021/jasms.0c00270.Search in Google Scholar PubMed
8. Plouffe, BD, Murthy, SK, Lewis, LH. Fundamentals and application of magnetic particles in cell isolation and enrichment: a review. Rep Prog Phys 2015;78:016601. https://doi.org/10.1088/0034-4885/78/1/016601.Search in Google Scholar PubMed PubMed Central
9. He, J, Huang, M, Wang, D, Zhang, Z, Li, G. Magnetic separation techniques in sample preparation for biological analysis: a review. J Pharm Biomed Anal 2014;101:84–101. https://doi.org/10.1016/j.jpba.2014.04.017.Search in Google Scholar PubMed
10. Safarik, I, Safarikova, M. Magnetic techniques for the isolation and purification of proteins and peptides. Biomagn Res Technol 2004;2:1–17. https://doi.org/10.1186/1477-044X-2-7.Search in Google Scholar PubMed PubMed Central
11. Pohanka, M. Biosensors and bioanalytical devices based on magnetic particles: a review. Curr Med Chem 2021;28:2828–41. https://doi.org/10.2174/0929867327666200730213721.Search in Google Scholar PubMed
12. Fredolini, C, Byström, S, Pin, E, Edfors, F, Tamburro, D, Iglesias, MJ, et al.. Immunocapture strategies in translational proteomics. Expert Rev Proteomics 2016;13:83. https://doi.org/10.1586/14789450.2016.1111141.Search in Google Scholar PubMed PubMed Central
13. Halvorsen, TG, Reubsaet, L. Antibody based affinity capture LC-MS/MS in quantitative determination of proteins in biological matrices. TrAC Trends Anal Chem 2017;95:132–9. https://doi.org/10.1016/j.trac.2017.08.009.Search in Google Scholar
14. Fung, EN, Bryan, P, Kozhich, A. Techniques for quantitative LC-MS/MS analysis of protein therapeutics: advances in enzyme digestion and immunocapture. Bioanalysis 2016;8:847–56. https://doi.org/10.4155/bio.16.24.Search in Google Scholar PubMed
15. Schuster, O, Atiya-Nasagi, Y, Rosen, O, Zvi, A, Glinert, I, Shmuel, AB, et al.. Coupling immuno-magnetic capture with LC-MS/MS(MRM) as a sensitive, reliable, and specific assay for SARS-CoV-2 identification from clinical samples. Anal Bioanal Chem 2022;414:1949–62. https://doi.org/10.1007/s00216-021-03831-5.Search in Google Scholar PubMed PubMed Central
16. Pierce, CL, Williams, TL, Santana, WI, Levine, M, Chen, LM, Cooper, HC, et al.. Immunocapture isotope dilution mass spectrometry in response to a pandemic influenza threat. Vaccine 2017;35:5011–8. https://doi.org/10.1016/j.vaccine.2017.07.049.Search in Google Scholar PubMed
17. Sleumer, B, Van Faassen, M, Vos, MJ, Bischoff, R, Kema, IP, van de Merbel, NC. Selective quantification of the 22-kDa isoform of human growth hormone 1 in serum and plasma by immunocapture and LC–MS/MS. Anal Bioanal Chem 2022;414:6187–200. https://doi.org/10.1007/s00216-022-04188-z.Search in Google Scholar PubMed PubMed Central
18. Huang, X, Xu, X, Partridge, MA, Chen, J, Koehler-Stec, E, Sumner, G, et al.. Isotyping and semi-quantitation of monkey anti-drug antibodies by immunocapture liquid chromatography-mass spectrometry. AAPS J 2021;23:1–11. https://doi.org/10.1208/s12248-020-00538-w.Search in Google Scholar PubMed PubMed Central
19. Zhao, Y, Gu, H, Postelnek, J, DeMichele, M, Yuan, L, Zhang, YJ, et al.. Fit-for-purpose protein biomarker assay validation strategies using hybrid immunocapture-liquid chromatography-tandem-mass spectrometry platform: quantitative analysis of total soluble cluster of differentiation 73. Anal Chim Acta 2020;1126:144–53. https://doi.org/10.1016/j.aca.2020.06.023.Search in Google Scholar PubMed
20. Whiteaker, JR, Zhao, L, Zhang, HY, Feng, LC, Piening, BD, Anderson, L, et al.. Antibody-based enrichment of peptides on magnetic beads for mass-spectrometry-based quantification of serum biomarkers. Anal Biochem 2007;362:44–54. https://doi.org/10.1016/j.ab.2006.12.023.Search in Google Scholar PubMed PubMed Central
21. Duggan, J, Ren, B, Mao, Y, Chen, LZ, Philip, E. LC-MS quantification of protein drugs: validating protein LC-MS methods with predigestion immunocapture. Bioanalysis 2016;8:1951–64. https://doi.org/10.4155/bio-2016-0137.Search in Google Scholar PubMed
22. Winther, B, Nordlund, M, Paus, E, Reubsaet, L, Halvorsen, TG. Immuno-capture as ultimate sample cleanup in LC-MS/MS determination of the early stage biomarker ProGRP. J Separ Sci 2009;32:2937–43. https://doi.org/10.1002/jssc.200900233.Search in Google Scholar PubMed
23. Kushnir, MM, Rockwood, AL, Strathmann, FG, Frank, EL, Straseski, JA, Meikle, AW. LC-MS/MS measurement of parathyroid hormone–related peptide. Clin Chem 2016;62:218–26. https://doi.org/10.1373/clinchem.2015.244012.Search in Google Scholar PubMed
24. Kumar, V, Barnidge, DR, Chen, LS, Twentyman, JM, Cradic, KW, Grebe, SK, et al.. Quantification of serum 1–84 parathyroid hormone in patients with hyperparathyroidism by immunocapture in situ digestion liquid chromatography–tandem mass spectrometry. Clin Chem 2010;56:306–13. https://doi.org/10.1373/clinchem.2009.134643.Search in Google Scholar PubMed
25. Stoll, DR, Carr, PW. Two-dimensional liquid chromatography: a state of the art tutorial. Anal Chem 2017;89:519–31. https://doi.org/10.1021/acs.analchem.6b03506.Search in Google Scholar PubMed
26. Levernæs, MCS, Broughton, MN, Reubsaet, L, Halvorsen, TG. To elute or not to elute in immunocapture bottom-up LC-MS. J Chromatogr B Anal Technol Biomed Life Sci 2017;1055–1056:51–60. https://doi.org/10.1016/j.jchromb.2017.03.044.Search in Google Scholar PubMed
27. Zheng, N, Taylor, K, Gu, H, Santockyte, R, Wang, XT, McCarty, J, et al.. Antipeptide immunocapture with in-sample calibration curve strategy for sensitive and robust LC-MS/MS bioanalysis of clinical protein biomarkers in formalin-fixed paraffin-embedded tumor tissues. Anal Chem 2020;92:14713–22. https://doi.org/10.1021/acs.analchem.0c03271.Search in Google Scholar PubMed
28. Berg, D, Malinowsky, K, Reischauer, B, Wolff, C, Becker, KF. Use of formalin-fixed and paraffin-embedded tissues for diagnosis and therapy in routine clinical settings. Methods Mol Biol 2011;785:109–22. https://doi.org/10.1007/978-1-61779-286-1_8.Search in Google Scholar PubMed
29. Escobar, H, Kushnir, MM, Ray, JA, Merrell, MA, Gomez, G, Fietkau, R, et al.. Measurement of pancreatic polypeptide and its peptide variant in human serum and plasma by immunocapture-liquid-chromatography-tandem mass spectrometry ref intervals pract assay considerations. Biochem Physiol 2014;3:140.10.4172/2168-9652.1000140Search in Google Scholar
30. Ulmer, C, Kritmetapak, K, Singh, R, Vesper, H, Kumar, R. High-resolution mass spectrometry for the measurement of PTH and PTH fragments: insights into PTH physiology and bioactivity. J Am Soc Nephrol 2022;38:1448–58. https://doi.org/10.1681/asn.2022010036.Search in Google Scholar PubMed PubMed Central
31. Kritmetapak, K, Losbanos, LA, Hines, JM, O’Grady, KL, Ulmer, CZ, Vesper, HW, et al.. Chemical characterization and quantification of circulating intact PTH and PTH fragments by high-resolution mass spectrometry in chronic renal failure. Clin Chem 2021;67:843–53. https://doi.org/10.1093/clinchem/hvab013.Search in Google Scholar PubMed PubMed Central
32. Budelier, MM, He, Y, Barthelemy, NR, Jiang, H, Li, Y, Park, E, et al.. A map of neurofilament light chain species in brain and cerebrospinal fluid and alterations in Alzheimer’s disease. Brain Commun 2022;4:fcac045. https://doi.org/10.1093/braincomms/fcac045.Search in Google Scholar PubMed PubMed Central
33. Liu, H, Zhang, N, Wan, D, Cui, M, Liu, Z, Liu, S. Mass spectrometry-based analysis of glycoproteins and its clinical applications in cancer biomarker discovery. Clin Proteonomics 2014;11:1–9. https://doi.org/10.1186/1559-0275-11-14.Search in Google Scholar PubMed PubMed Central
34. Chen, R, Tan, Y, Wang, M, Wang, F, Yao, Z, Dong, L, et al.. Development of glycoprotein capture-based label-free method for the high-throughput screening of differential glycoproteins in hepatocellular carcinoma. Mol Cell Proteomics 2011;10:M110.006445. https://doi.org/10.1074/mcp.m110.006445.Search in Google Scholar PubMed PubMed Central
35. DelGuidice, CE, Ismaiel, OA, Mylott, WR, Yuan, M, Halquist, MS. Intact quantitative bioanalytical method development and fit-for-purpose validation of a monoclonal antibody and its related fab fragment in human vitreous and aqueous humor using LC-HRMS. Anal Bioanal Chem 2022;414:4189–202. https://doi.org/10.1007/s00216-022-04071-x.Search in Google Scholar PubMed
36. Jin, W, Burton, L, Moore, I. LC-HRMS quantitation of intact antibody drug conjugate trastuzumab emtansine from rat plasma. Bioanalysis 2018;10:851–62. https://doi.org/10.4155/bio-2018-0003.Search in Google Scholar PubMed
37. Ravela, S, Valmu, L, Domanskyy, M, Koistinen, H, Kylänpää, L, Lindström, O, et al.. An immunocapture-LC-MS-based assay for serum SPINK1 allows simultaneous quantification and detection of SPINK1 variants. Anal Bioanal Chem 2018;410:1679–88. https://doi.org/10.1007/s00216-017-0803-y.Search in Google Scholar PubMed
38. Thibault, P, Perreault, C. Immunopeptidomics: reading the immune signal that defines self from nonself. Mol Cell Proteomics 2022;21:100234. https://doi.org/10.1016/j.mcpro.2022.100234.Search in Google Scholar PubMed PubMed Central
39. Chong, C, Coukos, G, Bassani-Sternberg, M. Identification of tumor antigens with immunopeptidomics. Nat Biotechnol 2021;40:175–88. https://doi.org/10.1038/s41587-021-01038-8.Search in Google Scholar PubMed
40. Kovalchik, KA, Wessling, L, Saab, F, Ma, Q, Despault, J, Kubiniok, P, et al.. Immunopeptidomics for dummies: detailed experimental protocols and rapid, user-friendly visualization of MHC I and II ligand datasets with MhcVizPipe. bioRxiv 2020;2020.11.02.360958.10.1101/2020.11.02.360958Search in Google Scholar
41. Xin, L, Qiao, R, Chen, X, Tran, H, Pan, S, Rabinoviz, S, et al.. A streamlined platform for analyzing tera-scale DDA and DIA mass spectrometry data enables highly sensitive immunopeptidomics. Nat Commun 2022;13:1–9. https://doi.org/10.1038/s41467-022-30867-7.Search in Google Scholar PubMed PubMed Central
42. Zhao, Y, Liu, G, Yuan, X, Gan, J, Peterson, JE, Shen, JX. Strategy for the quantitation of a protein conjugate via hybrid immunocapture-liquid chromatography with sequential HRMS and SRM-based LC-MS/MS analyses. Anal Chem 2017;89:5144–51. https://doi.org/10.1021/acs.analchem.7b00926.Search in Google Scholar PubMed
43. Dong, M. New HPLC, MS, and CDS products introduced in 2021–2022: a brief review. LCGC North Am 2022;40:165–8, 170–3. https://doi.org/10.56530/lcgc.na.jw6767l7.Search in Google Scholar
44. Sugimoto, H, Wei, D, Dong, L, Ghosh, D, Chen, S, Qian, MG. Perspectives on potentiating immunocapture-LC-MS for the bioanalysis of biotherapeutics and biomarkers. Bioanalysis 2018;10:1679–90. https://doi.org/10.4155/bio-2018-0205.Search in Google Scholar PubMed
45. Kellie, JF, Kehler, JR, Mencken, TJ, Snell, RJ, Hottenstein, CS. A whole-molecule immunocapture LC-MS approach for the in vivo quantitation of biotherapeutics. Bioanalysis 2016;8:2103–14. https://doi.org/10.4155/bio-2016-0180.Search in Google Scholar PubMed
46. Millet, A, Khoudour, N, Lebert, D, Machon, C, Terrier, B, Blanchet, B, et al.. Development, validation, and comparison of two mass spectrometry methods (LC-MS/HRMS and LC-MS/MS) for the quantification of rituximab in human plasma. Molecules 2021;26:1383. https://doi.org/10.3390/molecules26051383.Search in Google Scholar PubMed PubMed Central
47. Zubarev, RA, Makarov, A. Orbitrap mass spectrometry. Anal Chem 2013;85:5288–96. https://doi.org/10.1021/ac4001223.Search in Google Scholar PubMed
48. Jian, W, Kang, L, Burton, L, Weng, N. A workflow for absolute quantitation of large therapeutic proteins in biological samples at intact level using LC-HRMS. Bioanalysis 2016;8:1679–91. https://doi.org/10.4155/bio-2016-0096.Search in Google Scholar PubMed
49. Kellie, JF. Intact protein LC–MS for pharmacokinetics. Int J Pharmacokinet 2020;4:IPK05. https://doi.org/104155/ipk-2020-0004.10.4155/ipk-2020-0004Search in Google Scholar
50. Muccio, S, Tavernier, A, Rouchon, MC, Roccon, A, Dai, S, Finn, G, et al.. Validated method based on immunocapture and liquid chromatography coupled to high-resolution mass spectrometry to eliminate isatuximab interference with M-protein measurement in serum. Anal Chem 2021;93:15236–42. https://doi.org/10.1021/acs.analchem.1c03410.Search in Google Scholar PubMed
51. Finn, G, Macé, S, Campana, F, Le-Guennec, S, Muccio, S, Tavernier, A, et al.. Evaluating Isatuximab interference with monoclonal protein detection by immuno-capture and liquid chromatography coupled to high resolution mass spectrometry in the pivotal phase 3 multiple myeloma trial, icaria-MM. Blood 2019;134(1 Suppl):3143. https://doi.org/10.1182/blood-2019-129963.Search in Google Scholar
52. Himmelsbach, M. 10 years of MS instrumental developments – impact on LC–MS/MS in clinical chemistry. J Chromatogr B 2012;883–884:3–17. https://doi.org/10.1016/j.jchromb.2011.11.038.Search in Google Scholar PubMed
53. Ramanathan, R, Jemal, M, Ramagiri, S, Xia, YQ, Humpreys, WG, Olah, T, et al.. It is time for a paradigm shift in drug discovery bioanalysis: from SRM to HRMS. J Mass Spectrom 2011;46:595–601. https://doi.org/10.1002/jms.1921.Search in Google Scholar PubMed
54. Pang, Z, Zhou, G, Ewald, J, Chang, L, Hacariz, O, Basu, N, et al.. Using MetaboAnalyst 5.0 for LC–HRMS spectra processing, multi-omics integration and covariate adjustment of global metabolomics data. Nat Protoc 2022;17:1735–61. https://doi.org/10.1038/s41596-022-00710-w.Search in Google Scholar PubMed
55. Gutknecht, MF, Kaku, H, Rothstein, TL. Microparticle immunocapture assay for quantitation of protein multimer amount and size. Cell Rep Methods 2022;2:100214. https://doi.org/10.1016/j.crmeth.2022.100214.Search in Google Scholar PubMed PubMed Central
56. Bults, P, Sonesson, A, Knutsson, M, Bischoff, R, van de Merbel, NC. Intact protein quantification in biological samples by liquid chromatography – high-resolution mass spectrometry: somatropin in rat plasma. J Chromatogr B Anal Technol Biomed Life Sci 2020;15:1144. https://doi.org/10.1016/j.jchromb.2020.122079.Search in Google Scholar PubMed
57. Wang, L, Dodge, RW, Ji, QC. The future of immunocapture-capillary electrophoresis-high resolution mass spectrometry. Bioanalysis 2017;9:1627–31. https://doi.org/10.4155/bio-2017-4977.Search in Google Scholar PubMed
© 2023 Walter de Gruyter GmbH, Berlin/Boston
