Abstract
This chapter introduces bioactivity and bioaffinity terms in relation to mixture profiling and gives the significance of bioactivity and/or bioaffinity profiling of biologically active mixtures in general, and for bioactive mixtures in drug discovery research in particular. Further, the chapter gives an overview of the common and less common analytical approaches for bioactivity profiling of bioactive mixtures. Special focus is put on bioassay-guided fractionation as the standard technique employed (in identification and purification of bioactive molecules from a bioactive mixture), and on state-of-the-art post-column bioactivity profiling approaches, also providing examples and limitations of these analytical methods. On-column and pre-column bioactivity profiling analytics is also discussed. Examples of bioactive molecules identified and purified from different natural products are given with emphasis on molecules isolated from animal venoms. Finally, this chapter briefly discusses the importance of bioactivity profiling of metabolic mixtures in drug discovery.
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References
Liu B, Li S, Hu J (2004) Technological advances in high-throughput screening. Am J Pharmacogenomics 4(4):263–276
Weller MG (2012) A unifying review of bioassay-guided fractionation, effect-directed analysis and related techniques. Sensors (Basel) 12(7):9181–9209
Jonker W et al (2015) Methodologies for effect-directed analysis: environmental applications, food analysis, and drug discovery, in analyzing biomolecular interactions by mass spectrometry. Wiley-VCH, Weinheim, pp 109–163
Chen W et al (2015) Fasxiator, a novel factor XIa inhibitor from snake venom, and its site-specific mutagenesis to improve potency and selectivity. J Thromb Haemost 13(2):248–261
Graudins A et al (2012) Cloning and activity of a novel alpha-latrotoxin from red-back spider venom. Biochem Pharmacol 83(1):170–183
Feng J, Yang XW, Wang RF (2011) Bio-assay guided isolation and identification of alpha-glucosidase inhibitors from the leaves of Aquilaria sinensis. Phytochemistry 72(2–3):242–247
Crawford AD et al (2011) Zebrafish bioassay-guided natural product discovery: isolation of angiogenesis inhibitors from East African medicinal plants. PLoS One 6(2):e14694
Eng Kiat Loo A, Huang D (2007) Assay-guided fractionation study of α-amylase inhibitors from Garcinia mangostana pericarp. J Agri Food Chem 55(24):9805–9810
Su B-N et al (2002) Activity-guided fractionation of the seeds of Ziziphus jujuba using a cyclooxygenase-2 inhibitory assay. Planta Medica 68(12):1125–1128
Scher JM et al (2004) Bioactivity guided isolation of antifungal compounds from the liverwort Bazzania trilobata (L.) SF Gray. Phytochemistry 65(18):2583–2588
Ho CC, Kumaran A, Hwang LS (2009) Bio-assay guided isolation and identification of anti-Alzheimer active compounds from the root of Angelica sinensis. Food Chem 114(1):246–252
Awad R et al (2009) Bioassay-guided fractionation of lemon balm (Melissa officinalis L.) using an in vitro measure of GABA transaminase activity. Phytotherapy Res 23(8):1075–1081
Wu H et al (2013) Recent developments in qualitative and quantitative analysis of phytochemical constituents and their metabolites using liquid chromatography-mass spectrometry. J Pharm Biomed Anal 72:267–291
Bakker RA et al (2004) Constitutively active Gq/11-coupled receptors enable signaling by co-expressed G(i/o)-coupled receptors. J Biol Chem 279(7):5152–5161
Ohi N et al (1986) Semisynthetic Beta-Lactam antibiotics. 1. Synthesis and antibacterial activity of new ureidopenicillin derivatives having catechol moieties. J Antibiotics 39(2):230–241
Elander RP (2003) Industrial production of beta-lactam antibiotics. Appl Microbiol Biotechnol 61(5–6):385–392
Kondo S, Hotta K (1999) Semisynthetic aminoglycoside antibiotics: development and enzymatic modifications. J Infect Chemother 5(1):1–9
Manzoni M, Rollini N (2002) Biosynthesis and biotechnological production of statins by filamentous fungi and application of these cholesterol-lowering drugs. Appl Microbiol Biotechnol 58(5):555–564
Jonker N et al (2011) Recent developments in protein-ligand affinity mass spectrometry. Anal Bioanal Chem 399(8):2669–2681
Kool J et al (2011) Advances in mass spectrometry-based post-column bioaffinity profiling of mixtures. Anal Bioanal Chem 399(8):2655–2668
Kinawi A, Teller C (1979) Determination of drug-albumin binding in buffered bovine serum-albumin solutions applying a modified ultrafiltration process. Arzneimittel-Forschung/Drug Res 29–2(10):1495–1500
Zlotos G et al (1998) Determination of protein binding of gyrase inhibitors by means of continuous ultrafiltration. Pergamon-Elsevier Science, Amsterdam
Comess KM et al (2006) An ultraefficient affinity-based high-throughout screening process: application to bacterial cell wall biosynthesis enzyme MurF. J Biomol Screen 11(7):743–754
Li HL et al (2009) Screening and structural characterization of alpha-Glucosidase inhibitors from hawthorn leaf flavonoids extract by ultrafiltration LC-DAD-MSn and SORI-CID FTICR MS. J Am Soc Mass Spectrom 20(8):1496–1503
Ionita IA, Akhlaghi F (2010) Quantification of unbound prednisolone, prednisone, cortisol and cortisone in human plasma by ultrafiltration and direct injection into liquid chromatography tandem mass spectrometry. Ann Clin Biochem 47(Pt 4):350–357
Mulabagal V, Calderon AI (2010) Development of an ultrafiltration-liquid chromatography/mass spectrometry (UF-LC/MS) based ligand-binding assay and an LC/MS based functional assay for Mycobacterium tuberculosis shikimate kinase. Anal Chem 82(9):3616–3621
vanBreemen RB et al (1997) Pulsed ultrafiltration mass spectrometry: a new method for screening combinatorial libraries. Anal Chem 69(11):2159–2164
Geun Shin Y, Bolton JL, van Breemen RB (2002) Screening drugs for metabolic stability using pulsed ultrafiltration mass spectrometry. Comb Chem High Throughput Screen 5(1):59–64
Liu DT et al (2007) Screening for ligands of human retinoid X receptor-alpha. Using ultrafiltration mass spectrometry. Anal Chem 79(24):9398–9402
Cao H et al (2010) Discovery of cyclooxygenase inhibitors from medicinal plants used to treat inflammation. Pharmacol Res 61(6):519–524
Annis DA et al (2004) A general technique to rank protein-ligand binding affinities and determine allosteric versus direct binding site competition in compound mixtures. J Am Chem Soc 126(47):15495–15503
Derks RJE et al (2006) SEC-MS as an approach to isolate and directly identifying small molecular GPCR-ligands from complex mixtures without labeling. Chromatographia 64(7–8):379–385
Annis DA et al (2004) An affinity selection-mass spectrometry method for the identification of small molecule ligands from self-encoded combinatorial libraries—discovery of a novel antagonist of E-coli dihydrofolate reductase. Int J Mass Spectrom 238(2):77–83
Annis DA et al (2009) Inhibitors of the lipid phosphatase SHIP2 discovered by high-throughput affinity selection-mass spectrometry screening of combinatorial libraries. Comb Chem High Throughput Screen 12(8):760–771
Whitehurst CE et al (2006) Discovery and characterization of orthosteric and allosteric muscarinic M-2 acetylcholine receptor ligands by affinity selection-mass spectrometry. J Biomol Screen 11(2):194–207
Whitehurst CE, Annis DA (2008) Affinity selection-mass spectrometry and its emerging application to the high throughput screening of G protein-coupled receptors. Comb Chem High Throughput Screen 11(6):427–438
Jonker N et al (2008) Screening of protein-ligand interactions using dynamic protein-affinity chromatography solid-phase extraction-liquid chromatography-mass spectrometry. J Chromatogr A 1205(1–2):71–77
Hu F, Deng C, Zhang X (2008) Development of high performance liquid chromatography with immobilized enzyme onto magnetic nanospheres for screening enzyme inhibitor. J Chromatogr B 871(1):67–71
Marsza MP et al (2008) Ligand and protein fishing with heat shock protein 90 coated magnetic beads. Anal Chem 80(19):7571–7575
Jonker N et al (2009) Online magnetic bead dynamic protein-affinity selection coupled to LC-MS for the screening of pharmacologically active compounds. Anal Chem 81(11):4263–4270
Pochet L et al (2011) Online magnetic bead based dynamic protein affinity selection coupled to LC-MS for the screening of acetylcholine binding protein ligands. J Chromatogr B Analyt Technol Biomed Life Sci 879(20):1781–1788
Höfner G, Wanner KT (2015) MS binding assays. In: Analyzing biomolecular interactions by mass spectrometry. Wiley-VCH, Weinheim, pp 165–198
Zepperitz C, Hofner G, Wanner KT (2006) MS-binding assays: kinetic, saturation, and competitive experiments based on quantitation of bound marker as exemplified by the GABA transporter mGAT1. ChemMedChem 1(2):208–217
Singh NS, Jiang Z, Moaddel R (2015) Frontal and zonal affinity chromatography coupled to mass spectrometry. In: Analyzing biomolecular interactions by mass spectrometry. Wiley-VCH, Weinheim, pp 241–270
Loun B, Hage DS (1992) Characterization of thyroxine-albumin binding using high-performance affinity chromatography. I. Interactions at the warfarin and indole sites of albumin. J Chromatogr 579(2):225–235
Chan NW et al (2003) Frontal affinity chromatography-mass spectrometry assay technology for multiple stages of drug discovery: applications of a chromatographic biosensor. Anal Biochem 319(1):1–12
Moaddel R et al (2005) Enantioselective binding to the human organic cation transporter-1 (hOCT1) determined using an immobilized hOCT1 liquid chromatographic stationary phase. Chirality 17(8):501–506
Calleri E et al (2010) Frontal affinity chromatography-mass spectrometry useful for characterization of new ligands for GPR17 receptor. J Med Chem 53(9):3489–3501
Pharmaceuticals N. Tegretol (carbamazepine) extended-release tablets prescribing information. In N. Pharmaceuticals (ed). 2003
Haselberg R et al (2014) Capillary electrophoresis-based assessment of nanobody affinity and purity. Anal Chim Acta 818:1–6
Medina-Casanellas S et al (2012) Preparation and evaluation of an immunoaffinity sorbent for the analysis of opioid peptides by on-line immunoaffinity solid-phase extraction capillary electrophoresis-mass spectrometry. Anal Chim Acta 717:134–142
Haselberg R, Somsen GW (2015) Online affinity assessment and immunoaffinity sample pretreatment in capillary electrophoresis–mass spectrometry, in analyzing biomolecular interactions by mass spectrometry. Wiley-VCH, Weinheim, pp 271–298
Nijmeijer S et al (2012) Development of a profiling strategy for metabolic mixtures by combining chromatography and mass spectrometry with cell-based GPCR signaling. J Biomol Screen 17(10):1329–1338
Giera M et al (2009) Microfractionation revisited: a 1536 well high resolution screening assay. Anal Chem 81(13):5460–5466
Falck D et al (2013) Development of on-line liquid chromatography-biochemical detection for soluble epoxide hydrolase inhibitors in mixtures. Chromatographia 76(1–2):13–21
Oosterkamp AJ et al (1997) Theoretical concepts of on-line liquid chromatographic- biochemical detection systems II. Detection systems based on labelled affinity proteins. J Chromatogr A 787(1–2):37–46
Oosterkamp AJ et al (1997) Theoretical concepts of on-line liquid chromatographic-biochemical detection systems. I. Detection systems based on labelled ligands. J Chromatogr A 787(1–2):27–35
Marques LA et al (2010) Production and on-line acetylcholinesterase bioactivity profiling of chemical and biological degradation products of tacrine. J Pharm Biomed Anal 53(3):609–616
Kool J et al (2007) Cytochrome P450 bio-affinity detection coupled to gradient HPLC: on-line screening of affinities to cytochrome P4501A2 and 2D6. J Chromatogr B 858(1):49–58
Falck D et al (2010) Development of an online p38alpha mitogen-activated protein kinase binding assay and integration of LC-HR-MS. Anal Bioanal Chem 398(4):1771–1780
Kool J et al (2010) Online fluorescence enhancement assay for the acetylcholine binding protein with parallel mass spectrometric identification. J Med Chem 53(12):4720–4730
de Vlieger JS et al (2010) Determination and identification of estrogenic compounds generated with biosynthetic enzymes using hyphenated screening assays, high resolution mass spectrometry and off-line NMR. J Chromatogr B Analyt Technol Biomed Life Sci 878(7–8):667–674
Schenk T et al (2003) A generic assay for phosphate-consuming or-releasing enzymes coupled on-line to liquid chromatography for lead finding in natural products. Anal Biochem 316(1):118–126
Hogenboom A et al (2001) Continuous-flow, on-line monitoring of biospecific interactions using electrospray mass spectrometry. Anal Chem 73(16):3816–3823
Heus F et al (2010) Development of a microfluidic confocal fluorescence detection system for the hyphenation of nano-LC to on-line biochemical assays. Anal Bioanal Chem 398(7–8):3023–3032
Otvos RA et al (2013) Analytical workflow for rapid screening and purification of bioactives from venom proteomes. Toxicon 76:270–281
Heus F et al (2014) Miniaturized bioaffinity assessment coupled to mass spectrometry for guided purification of bioactives from toad and cone snail. Biology 3(1):139–156
Heus F et al (2013) An efficient analytical platform for on-line microfluidic profiling of neuroactive snake venoms towards nicotinic receptor affinity. Toxicon 61:112–124
Giera M et al (2010) Structural elucidation of biologically active neomycin N-octyl derivatives in a regioisomeric mixture by means of liquid chromatography/ion trap time-of-flight mass spectrometry. Rapid Commun Mass Spectrom 24(10):1439–1446
Kool J et al (2012) High-resolution metabolic profiling towards G protein-coupled receptors: rapid and comprehensive screening of histamine H(4) receptor ligands. J Chromatogr A 1259:213–220
Mladic M et al (2016) At-line nanofractionation with parallel mass spectrometry and bioactivity assessment for the rapid screening of thrombin and factor Xa inhibitors in snake venoms. Toxicon 110:79–89
Mladic M et al (2017) Rapid screening and identification of ACE inhibitors in snake venoms using at-line nanofractionation LC-MS. Anal Bioanal Chem 409(25):5987–5997
Still KS et al (2017) Multipurpose HTS coagulation analysis: assay development and assessment of coagulopathic snake venoms. Toxins (Basel) 9(12):382
Bordon KC et al (2012) Isolation, enzymatic characterization and antiedematogenic activity of the first reported rattlesnake hyaluronidase from Crotalus durissus terrificus venom. Biochimie 94(12):2740–2748
Wiezel GA et al (2015) Identification of hyaluronidase and phospholipase B in Lachesis muta rhombeata venom. Toxicon 107(Pt B):359–368
Babaie M et al (2013) Isolation and partial purification of anticoagulant fractions from the venom of the Iranian snake Echis carinatus. Acta Biochim Pol 60(1):17–20
Menaldo DL et al (2015) Purification procedure for the isolation of a P-I metalloprotease and an acidic phospholipase A2 from Bothrops atrox snake venom. J Venom Anim Toxins Incl Trop Dis 21:28
Osipov AV et al (2017) New paradoxical three-finger toxin from the cobra Naja kaouthia venom: Isolation and characterization. Dokl Biochem Biophys 475(1):264–266
Teixeira TL et al (2016) Isolation, characterization and screening of the in vitro cytotoxic activity of a novel L-amino acid oxidase (LAAOcdt) from Crotalus durissus terrificus venom on human cancer cell lines. Toxicon 119:203–217
Rodriguez-Acosta A et al (2016) Biological and biochemical characterization of venom from the broad-banded copperhead (Agkistrodon contortrix laticinctus): isolation of two new dimeric disintegrins. Anim Biol Leiden Neth 66(2):173–187
Fucase TM et al (2017) Isolation and biochemical characterization of bradykinin-potentiating peptides from Bitis gabonica rhinoceros. J Venom Anim Toxins Incl Trop Dis 23:33
King GF (2011) Venoms as a platform for human drugs: translating toxins into therapeutics. Expert Opin Biol Ther 11(11):1469–1484
Ferreira SH, Bartelt DC, Greene LJ (1970) Isolation of bradykinin-potentiating peptides from Bothrops jararaca venom. Biochemistry 9(13):2583–2593
Ondetti MA et al (1971) Angiotensin-converting enzyme inhibitors from the venom of Bothrops jararaca. Isolation, elucidation of structure, and synthesis. Biochemistry 10(22):4033–4039
Smith CG, Vane JR (2003) The discovery of captopril. FASEB J 17(8):788–789
Scarborough RM (1999) Development of eptifibatide. Am Heart J 138(6 Pt 1):1093–1104
Cook JJ et al (1999) Tirofiban (Aggrastat (R)). Cardiovasc Drug Rev 17(3):199–224
Earl ST et al (2012) Drug development from Australian elapid snake venoms and the Venomics pipeline of candidates for haemostasis: Textilinin-1 (Q8008), Haempatch (Q8009) and CoVase (V0801). Toxicon 59(4):456–463
Koh CY, Kini RM (2012) From snake venom toxins to therapeutics–cardiovascular examples. Toxicon 59(4):497–506
Vink S et al (2012) Natriuretic peptide drug leads from snake venom. Toxicon 59(4):434–445
Diochot S et al (2012) Black mamba venom peptides target acid-sensing ion channels to abolish pain. Nature 490(7421):552–555
Pu XC, Wong PT, Gopalakrishnakone P (1995) A novel analgesic toxin (hannalgesin) from the venom of king cobra (Ophiophagus hannah). Toxicon 33(11):1425–1431
McCleary RJ, Kini RM (2013) Non-enzymatic proteins from snake venoms: a gold mine of pharmacological tools and drug leads. Toxicon 62:56–74
Kini RM, Doley R (2010) Structure, function and evolution of three-finger toxins: mini proteins with multiple targets. Toxicon 56(6):855–867
Lewis RJ et al (2012) Conus venom peptide pharmacology. Pharmacol Rev 64(2):259–298
Twede VD et al (2009) Neuroprotective and cardioprotective conopeptides: an emerging class of drug leads. Curr Opin Drug Discov Devel 12(2):231–239
Essack M, Bajic VB, Archer JA (2012) Conotoxins that confer therapeutic possibilities. Mar Drugs 10(6):1244–1265
Vetter I, Lewis RJ (2012) Therapeutic potential of cone snail venom peptides (conopeptides). Curr Top Med Chem 12(14):1546–1552
Miljanich GP (2004) Ziconotide: neuronal calcium channel blocker for treating severe chronic pain. Curr Med Chem 11(23):3029–3040
Stöcklin R. XEP-018: a new myorelaxant peptide lead compound from the venom of the cone snail Conus consors. Proceedings of 7th Annual Peptide Therapeutics Symposium, 2012. p. 32
Furman BL (2012) The development of Byetta (exenatide) from the venom of the Gila monster as an anti-diabetic agent. Toxicon 59(4):464–471
Turton M et al (1996) A role for glucagon-like peptide-1 in the central regulation of feeding. Nature 379(6560):69–72
Doyle ME, Egan JM (2000) Glucagon-like peptide-1. Recent Prog Horm Res 56:377–399
Klint JK et al (2012) Spider-venom peptides that target voltage-gated sodium channels: pharmacological tools and potential therapeutic leads. Toxicon 60(4):478–491
Mortari MR, Cunha AOS (2013) New perspectives in drug discovery using neuroactive molecules from the venom of arthropods. IntechOpen, London
Yang S et al (2013) Discovery of a selective NaV1. 7 inhibitor from centipede venom with analgesic efficacy exceeding morphine in rodent pain models. Proc Natl Acad Sci 110(43):17534–17539
Lewis RJ, Garcia ML (2003) Therapeutic potential of venom peptides. Nat Rev Drug Discov 2(10):790–802
Mathias NR, Hussain MA (2010) Non-invasive systemic drug delivery: developability considerations for alternate routes of administration. J Pharm Sci 99(1):1–20
Anderson S (2005) Making medicines: a brief history of pharmacy and pharmaceuticals. Pharmaceutical Press, London
Hamilton GR, Baskett TF (2000) In the arms of Morpheus the development of morphine for postoperative pain relief. Can J Anaesth 47(4):367–374
Sneader W (1996) Drug prototypes and their exploitation. Wiley, Chichester
Newman DJ, Cragg GM (2012) Natural products as sources of new drugs over the 30 years from 1981 to 2010. J Nat Prod 75(3):311–335
Koehn FE, Carter GT (2005) The evolving role of natural products in drug discovery. Nat Rev Drug Discov 4(3):206–220
Harvey AL (2008) Natural products in drug discovery. Drug Discov Today 13(19–20):894–901
Harvey AL (2014) Toxins and drug discovery. Toxicon 92:193–200
Cragg GM, Newman DJ (2013) Natural products: a continuing source of novel drug leads. Biochim Biophys Acta 1830(6):3670–3695
Chin YW et al (2006) Drug discovery from natural sources. AAPS J 8(2):E239–E253
Fabricant DS, Farnsworth NR (2001) The value of plants used in traditional medicine for drug discovery. Environ Health Perspect 109(Suppl 1):69–75
Hanks GW et al (2001) Morphine and alternative opioids in cancer pain: the EAPC recommendations. Br J Cancer 84(5):587–593
Furlan AD et al (2006) Opioids for chronic noncancer pain: a meta-analysis of effectiveness and side effects. CMAJ 174(11):1589–1594
Campbell TJ, Williams KM (2001) Therapeutic drug monitoring: antiarrhythmic drugs. Br J Clin Pharmacol 52:21s–34s
Barnes PJ (2006) Theophylline for COPD. Thorax 61(9):742–744
Barnes PJ (2001) Tiotropium bromide. Expert Opin Investig Drugs 10(4):733–740
Hansel TT, Barnes PJ (2002) Tiotropium bromide: a novel once-daily anticholinergic bronchodilator for the treatment of COPD. Drugs Today (Barc) 38(9):585–600
Elliott WJ, Ram CV (2011) Calcium channel blockers. J Clin Hypertens (Greenwich) 13(9):687–689
Bailey CJ, Day C (2004) Metformin: its botanical background. Pract Diab Int 21(3):115–117
Ruetsch YA, Boni T, Borgeat A (2001) From cocaine to ropivacaine: the history of local anesthetic drugs. Curr Top Med Chem 1(3):175–182
Raghavendra T (2002) Neuromuscular blocking drugs: discovery and development. J R Soc Med 95(7):363–367
Graziose R, Lila MA, Raskin I (2010) Merging traditional Chinese medicine with modern drug discovery technologies to find novel drugs and functional foods. Curr Drug Discov Technol 7(1):2
Gong X, Sucher NJ (1999) Stroke therapy in traditional Chinese medicine (TCM): prospects for drug discovery and development. Trends Pharmacol Sci 20(5):191–196
Hsiao W, Liu L (2010) The role of traditional Chinese herbal medicines in cancer therapy—from TCM theory to mechanistic insights. Planta Medica 76(11):1118
Lao L, Xu L, Xu S (2012) Traditional Chinese medicine. In: Integrative pediatric oncology. Springer, Beijing, pp 125–135
Scriabine A (1999) Discovery and development of major drugs currently in use. In: Pharmaceutical innovation: revolutionizing human health. Chemical Heritage Press, Philadelphia, pp 148–270
Page MG (2012) Beta-lactam antibiotics. In: Antibiotic discovery and development. Springer, New York, pp 79–117
Chopra I, Roberts M (2001) Tetracycline antibiotics: mode of action, applications, molecular biology, and epidemiology of bacterial resistance. Microbiol Mol Biol Rev 65(2):232–260; second page, table of contents
Forge A, Schacht J (2000) Aminoglycoside antibiotics. Audiol Neurootol 5(1):3–22
Omura S (2002) Macrolide antibiotics: chemistry, biology, and practice. Academic Press, Orlando
Berdy J (2005) Bioactive microbial metabolites. J Antibiot (Tokyo) 58(1):1–26
Colombo D, Ammirati E (2011) Cyclosporine in transplantation—a history of converging timelines. J Biol Regul Homeost Agents 25(4):493–504
Craik DJ et al (2013) The future of peptide-based drugs. Chem Biol Drug Des 81(1):136–147
Kola I, Landis J (2004) Can the pharmaceutical industry reduce attrition rates? Nat Rev Drug Discov 3(8):711–716
Cusack KP et al (2013) Emerging technologies for metabolite generation and structural diversification. Bioorg Med Chem Lett 23(20):5471–5483
Dhont M (2010) History of oral contraception. Eur J Contracept Reprod Health Care 15(S2):S12–S18
Lin JH, Lu AY (1997) Role of pharmacokinetics and metabolism in drug discovery and development. Pharmacol Rev 49(4):403–449
Mladic M et al (2015) At-line coupling of LC-MS to bioaffinity and selectivity assessment for metabolic profiling of ligands towards chemokine receptors CXCR1 and CXCR2. J Chromatogr B Analyt Technol Biomed Life Sci 1002:42–53
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Mladic, M., Niessen, W.M.A., Somsen, G.W., Kool, J. (2020). Analytics for Bioactivity Profiling of Complex Mixtures with a Focus on Venoms. In: Priel, A. (eds) Snake and Spider Toxins. Methods in Molecular Biology, vol 2068. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9845-6_2
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