Peptide Analysis: Critical Technology for Evaluating Delivery Approaches

  • Larry A. Sternson
  • Thomas R. Malefyt
Part of the NATO ASI Series book series (NSSA, volume 125)


Peptides and proteins are emerging as an increasingly important chemical class of drugs as they become more readily available through improvements in recombinant DNA technology and approaches to their chemical synthesis.


Drug Substance Capillary Zone Electrophoresis Peptide Drug Fluorescence Polarization Immunoassay Reporter Group 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Allen, R.C. and Maurer, H.R., 1974, “Electrophoresis and Isoelectric Focusing in Polyacrylamide Gel”, Walter de Gruyter, New York.Google Scholar
  2. Arbuthnot, J.P. and Beeley, J.A., 1975, “Isoelectric Focusing, Butterworths, London.Google Scholar
  3. Bachi, T., Gernard, W. and Yewdell, J.W., 1985, Monoclonal Antibodies Detect Different Forms of Influenza Virus Hemagglutinin During Viral Penetration and Biosynthesis, J.Virol., 55: 307-313.PubMedGoogle Scholar
  4. Barber, M., Bordoli, R.S., Garner, G.V., Gordon, D.B., Sedgwick, R.D., Tetler, L.W. and Tyler, A.N., 1981, Fast-Atom-Bombardment Mass Spectra of Enkephalins, Biochem.J., 197: 401.PubMedGoogle Scholar
  5. Blecka, L.J., 1983, Fluorescence Polarization Immunoassay: A Review of Methodology and Applications, Amer.Assoc.Clin.Chem., March 1.Google Scholar
  6. Bradford, M., 1976, A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein Dye Binding, Anal.Biochem., 72: 248.PubMedCrossRefGoogle Scholar
  7. Carr, S.A. and Roberts, G.D., 1986, Carbohydrate Mapping by Mass Spectrometry. A Novel Method for Identifying Attachment sites in Asn-linked Sugars in Glycoproteins, manuscript in preparation.Google Scholar
  8. Cinander, B., 1967, Antibodies to Biologically Active Molecules, in: “Proceedings of the Second Meeting of the Foundation of European Biochemical Societies, Vienna, Pergamon Press, Oxford.Google Scholar
  9. de Vlaminck, W., 1984, FDA’s Role in Approval and Regulation of Recombinant DNA Drugs, in: “Recombinant DNA Products: Insulin, Interferon and Growth Hormone”, A.P. Bollen, ed., CRC Press Inc., Boca Raton.Google Scholar
  10. Edman, P., 1950, Method for Determination of the Amino Acid Sequence in Peptides, Acta.Chem.Scand., 4: 283.CrossRefGoogle Scholar
  11. Engvall, E. and Perlman, P., 1971, Enzyme Linked Immunosorbant Assay (ELISA) Quantitative Assay of Immunoglobulin G, Immunochem., 8: 871.CrossRefGoogle Scholar
  12. Fareed, J., Walenga, H.L., Messmore, E.W. and Bermes, E.W., 1983, Synthetic Substrates in Hemostatic Testing, in: “CRC Critical Review in Laboratory Sciences”, CRC Press Inc., Boca Raton.Google Scholar
  13. Gordon, A.H., 1975, in: “Laboratory Techniques in Biochemistry and Molecular Biology”, T.W. Work and E. Work, eds., North Holland Publishing Co., Amsterdam.Google Scholar
  14. Hearn, M.T.W., 1983, High Performance Liquid Chromatography of Peptides, in: “HPLC Advances and Perspectives”, C. Horvath, ed., Academic Press, New York.Google Scholar
  15. Hearn, M.T.W, Regnier, F.E. and Wehr, C.T., 1983, HPLC of Peptides and Proteins”, Academic Press, New York.Google Scholar
  16. Imai, K., Toyo’oka, T. and Miyano, H., 1984, Fluorogenic Reagents for Primary and Secondary Amines and Thiols in High Performance Liquid Chromatography, Analyst, 109: 1365.CrossRefGoogle Scholar
  17. Jolley, M.E., 1981, Fluorescence Polarization Immunoassay for the Determination of Therapeutic Drug Levels in Human Plasma, J.Anal.Toxicol., 5: 236.PubMedCrossRefGoogle Scholar
  18. Jolley, M.E., Stroupe, S.D., Schwenzer, K.S., Wang, C.J., Lu-Steffes, M., Hill, H.D., Popelka, S.R., Holen, J.T. and Kelso, D.M., 1981b, Fluorescence Polarization Immunoassay III. An Automated System for Therapeutic Drug Determination, Clin.Chem., 27: 1575.PubMedGoogle Scholar
  19. Jorgenson, J.W. and Lukacs, K.D., 1983, Capillary Zone Electrophoresis, Science, 222: 266.PubMedCrossRefGoogle Scholar
  20. Jornvall, H., Pohl, G., Bergsdorf, W. and Wallen, P., 1983, Differential Proteolysis and Evidence for a Residue Exchange in Tissue-Plasminogen Activator Suggest Possible Association Between Two Types of Protein Microheterogeneity, FEBS Letters, 156: 47.PubMedCrossRefGoogle Scholar
  21. Kiss, I., Aurell, L., Pozsgay, M. and Elodi, P., 1985, Investigation on the Substrate Specificity of Human Plasmin Using Tripeptidyl p-nitroanilide Substrates, Biochem.Biophys.Res.Commun., 131: 928.PubMedCrossRefGoogle Scholar
  22. Kohler, G. and Milstein, C., 1975, Continuous Cultures of Fused Cells Secreting Antibody of Predefined Specificity, Nature, 256: 495.PubMedCrossRefGoogle Scholar
  23. Liversidge, G., Kinter, L. and Sternson, L., 1986, manuscript in preparation.Google Scholar
  24. Lowry, O.H. and Passonneau, J.V., 1972, “A Flexible System of Enzymatic Analysis, Academic Press, New York.Google Scholar
  25. Lowry, O.H., Rosebrough, N.J., Farr, A.L. and Randall, R.J., 1951, Protein Measurement with the Folin Phenol Reagent, J.Biol.Chem., 193: 265.PubMedGoogle Scholar
  26. Lukacs, K.D. and Jorgenson, J.W., 1981, Zone Electrophoresis in Open-Tublar Glass Capillaries: Preliminary Data on Performance, J.High Res.Chrom., 4: 230.CrossRefGoogle Scholar
  27. Messmore, H.L., Fareed, J., Kniffin, J., Squillaci, G. and Walenga, J.M., 1981, Synthetic Substrate Assays of the Coagulation Enzymes and Their Inhibitors. Comparison with clotting and Immunological Methods for Clinical and Experimental Usage, Ann.N.Y.Acad.Sci., 370: 787.CrossRefGoogle Scholar
  28. Ohno, Y., Kato, H., Iwanaga, S., Takada, K., Sakakibara, S. and Steuflo, J., 1981, A New Fluorogenic Peptide Substrate for Vitamin K Dependent Blood Coagulation Factor Bovine Protein C, J.Biochem., 90: 1387.PubMedGoogle Scholar
  29. Peterson, G., 1979, Review of the Folin Phenol Protein Quantitation Methoa of Lowry, Rosenbrough, Farr and Randall, Anal.Biochem., 100: 210.CrossRefGoogle Scholar
  30. Ranby, M., Bergsdorf, N., Pohl, G. and Wallen, P., 1982, Isolation of Two Variants of Native One-Chain Tissue-Plasminogen Activator, FEBS Letters, 146: 289.PubMedCrossRefGoogle Scholar
  31. Reghetti, P.G. and Drysdale, J.W., 1975, in: “Laboratory Techniques in Biochemistry and Molecular Biology”, T.S. Work and E. Work, eds., North Holland Publishing Co., Amsterdam.Google Scholar
  32. Rubenstein, K.E., Schneider, R.S. and Ullman, E.F., 1972, Homogeneous Enzyme Immunoassay. A New Immunochemical Technique, Biochem.Biophys Res.Commun., 47: 846.PubMedCrossRefGoogle Scholar
  33. Sanger, F., 1945, The Free Amino Groups of Insulin, Biochem.J., 39: 507.PubMedGoogle Scholar
  34. Schlabach, T.D., 1984, Post-Column Detection of Serum Proteins with the Biuret and Lowry Reactions, Anal.Biochem., 139: 309.PubMedCrossRefGoogle Scholar
  35. Schlabach, T.D. and Wehr, C.T., 1983, Fluorescent Techniques for the Selective Detection of Chromatographically Separated Peptides, in: “HPLC of Peptides and Proteins”, M.T.W. Hearn, F.E. Regnier and C.T. Wehr, ed., Academic Press, New York.Google Scholar
  36. Schonherr, O.T. and Houwink, E.H., 1984, Antibody Engineering, a Strategy for the Development of Monoclonal Antibodies, Ant.van Leewenhoek, 50: 597.CrossRefGoogle Scholar
  37. Schwarz, S., Berger, P. and Wick, G., 1985, Epitope-Selective, Monoclonal-Antibody-Based Immunoradiometric Assays of Predictable Specificity for Differential Measurement of Choriogonadotropin and Its Subunits, Clin.Chem., 31: 1322.PubMedGoogle Scholar
  38. Secher, D.S. and Burke, D.C., 1980, A Monoclonal Antibody for Large Scale Purification of Human leukocyte Interferon, Nature, 285: 446.PubMedCrossRefGoogle Scholar
  39. Smith, P.K., Krohn, R.I., Hermanson, G.T., Mallia, A.K., Gartner, F.H., Provenzano, M.D., Fujimoto, E.K., Groeke, N.M., Olson, B.J. and Klenk, D.C., 1985, Measurement of Protein Using Bicinchoninic Acid, Anal.Biochem., 150: 76.PubMedCrossRefGoogle Scholar
  40. Staehelin, T., Durrer, B., Schmidt, J., Takacs, B., Stocker, J., Miggiano, V., Stahli, C., Rubenstein, M., Levy, W.P., Hershberg, R. and Pestka, S., 1981, Production of Hybridomas Secreting Monoclonal Antibodies to the Human Leukocyte Interferons, Proc.Nat’l.Acad.Sci. U.S.A., 78: 1848.CrossRefGoogle Scholar
  41. Sternson, L.A., Higuchi, T., DeMontigny, P., Repta, A. and Stobaugh, J., 1985b, U.S. Patent, “Assaying Method for Primary Amines Using Aromatic Dialdehydes”, application filed.Google Scholar
  42. Sternson, L.A. and Malefyt, T.R., 1985a, Analytical Aspects of Drug Delivery: An Important and Often Overlooked Problem, in: “Directed Drug Delivery”, Humana Press, Clifton.Google Scholar
  43. Sternson, L.A., Repta, A.J. and Stobaugh, J.F., 1985c, Rational Design and Evaluation of Improved o-phthalaldehyde-like Fluorogenic Reagents, Anal.Biochem., 144: 233.PubMedCrossRefGoogle Scholar
  44. Svendsen, L.G., Fareed, J., Walenga, J.M. and Hoppensteadt, D., 1983, Newer Synthetic Peptide Substrates in Coagulation Testing: Some Practical Considerations for Automated Methods, Seminars in Thrombosis and Hemostasis, 9: 250.PubMedCrossRefGoogle Scholar
  45. Tang, J., Li, S., McGray, P. and Vecchio, A., 1984, Current Status of Activity Assays for Tissue-Plasminogen Activator, Ann.New York Acad.Sci., 434: 536.CrossRefGoogle Scholar
  46. Wallen, P., Pohl, G., Ranby, M. and Jornvall, H., 1983, Purification and Characterization of a Melanoma Cell Plasminogen Activator, Eur.J.Biochem., 132: 681.PubMedCrossRefGoogle Scholar
  47. Wilson, M.A., Kao, Y. and Miles, L., 1981, Direct Comparison of a Radioimmunoassay and an Immunoradiometric Technique in the Measurement of Human Growth Hormone, Nuc.Med.Commun., 2: 68.Google Scholar
  48. Witt, I., Tritschler, W. and Bablok, W., 1981a, Alpha 2-macroglobulin: Values in Serum and Plasma with a Chromogenic Substrate, J.Clin.Chem.Clin.Biochem., 19: 887.Google Scholar
  49. Witt, I., 1981b, New Methods for the Analysis of Coagulation Using Chromogenic Substrate, J.C1in.Chem.Clin.Biochem., 19: 877.Google Scholar
  50. Yewdell, J.W., Gernard, W. and Bachi, T., 1983, Monoclonal Hemagglutinin Antibodies Direct Irreversible Antigenic Alterations that Coincide with the Acid Activation of Influenza Virus A/PR/834-Mediated Hemolysis, J.Virol., 48: 239.PubMedGoogle Scholar
  51. Ying, S.Y., 1981, Radioimmunoassay of Peptide Hormones Using Killed S. Aureus as a Separation Agent, in: “Immunochemical Techniques, Methods in Enzymology”, J.J. Langone and H. VanVunakis, eds., Academic Press, New York.Google Scholar
  52. Zimmerman, C.L., Apella, E. and Pisano, J.J., 1977, Rapid Analysis of Amino Acid Phenylthiohydantoins by High Performance Liquid Chromatography, Anal.Biochem., 77: 569.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1986

Authors and Affiliations

  • Larry A. Sternson
    • 1
  • Thomas R. Malefyt
    • 1
  1. 1.Smith Kline & French LaboratoriesSwedelandUSA

Personalised recommendations