Biological and Biomedical Applications of Isoelectric Focusing

It has now been over a decade since isoelectric focusing became estab lished as a valuable addition to our arsenal of physical methods for separa tion of proteins and other amphoteric substances. The high resolving power of the technique and its unique ability to separate as well as concentrate have been amply demonstrated in several international symposia. The past few years have led to the wide acceptance of standardized methodologies for analytical and preparative purposes. These procedures afford rapid, reproducible separations that have led to a dramatic increase in our under standing of many areas of biological and biomedical research. Conse quently, we considered it desirable to review some of the important appli cations of the isoelectric focusing technique. To accomplish this, we have solicited the participation of noted authorities to highlight major progress in their fields of expertise. Thus, the material in this book will emphasize recent advances in knowledge rather than methodological aspects, except when special procedures are reviewed. Nicholas Catsimpoolas James W. Drysdale vii Contents Chapter J Isoelectric Focusing of Human Saliva, Cerebrospinal Fluid, and Urine Josie A. Beeley I. Introduction I II. Isoelectric Focusing of Saliva . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 A. Gel Rods and Thin Layer Gels . . . . . . . . . . . . . . . . . . . . . . . . 2 B. Liquid Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 C. a-Amylases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 D. Immunoglobulins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 E. Blood Group Specific Substances . . . . . . . . . . . . . . . . . . . . . 11 F. Vitamin B12 Binding Proteins (Cobalophilins) . . . . . . . . . . . 12 G. Other Glycoproteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 H. Lysozyme . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 I. Specific Salivary Proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 J. Dental Plaque . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 K. Other Proteins . . . . . . . . . . . . . . . , . . . . . . . . . . . . . . . . . . . . . . 16 III. Isoelectric Focusing of Cerebrospinal Fluid . . . . . . . . . . . . . . . .

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It has now been over a decade since isoelectric focusing became estab­ lished as a valuable addition to our arsenal of physical methods for separa­ tion of proteins and other amphoteric substances. The high resolving power of the technique and its unique ability to separate as well as concentrate have been amply demonstrated in several international symposia. The past few years have led to the wide acceptance of standardized methodologies for analytical and preparative purposes. These procedures afford rapid, reproducible separations that have led to a dramatic increase in our under­ standing of many areas of biological and biomedical research. Conse­ quently, we considered it desirable to review some of the important appli­ cations of the isoelectric focusing technique. To accomplish this, we have solicited the participation of noted authorities to highlight major progress in their fields of expertise. Thus, the material in this book will emphasize recent advances in knowledge rather than methodological aspects, except when special procedures are reviewed. Nicholas Catsimpoolas James W. Drysdale vii Contents Chapter J Isoelectric Focusing of Human Saliva, Cerebrospinal Fluid, and Urine Josie A. Beeley I. Introduction I II. Isoelectric Focusing of Saliva . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 A. Gel Rods and Thin Layer Gels . . . . . . . . . . . . . . . . . . . . . . . . 2 B. Liquid Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 C. a-Amylases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 D. Immunoglobulins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 E. Blood Group Specific Substances . . . . . . . . . . . . . . . . . . . . . 11 F. Vitamin B12 Binding Proteins (Cobalophilins) . . . . . . . . . . . 12 G. Other Glycoproteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 H. Lysozyme . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 I. Specific Salivary Proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 J. Dental Plaque . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 K. Other Proteins . . . . . . . . . . . . . . . , . . . . . . . . . . . . . . . . . . . . . . 16 III. Isoelectric Focusing of Cerebrospinal Fluid . . . . . . . . . . . . . . . .

Contenu
1 Isoelectric Focusing of Human Saliva, Cerebrospinal Fluid, and Urine.- I. Introduction.- II. Isoelectric Focusing of Saliva.- III. Isoelectric Focusing of Cerebrospinal Fluid.- IV. Isoclectdc Focusing of Urine.- V. Conclusions.- VI.References.- 2 The Use of Gel in the Analysis of Hemoglobins.- I. Introduction.- II. Practical Applications.- III. Experimental Applications.- IV. References.- 3 Isoelectric Focusing of Allergens.- I. Introduction.- II. Methods.- III. Isoelectric Focusing of Allergens.- IV. Applications.- V. General Comments.- VI. References.- 4 Isoelectric Focusing of Muscle Proteins.- I. Introduction.- II. Technical Considerations.- III. Specific Applications.- IV. Conclusions and Predictions.- V. References.- 5 Studies of the Structure and Synthesis of Immunoglobulins by Isoelectric Focusing.- I. Introduction.- II. Methods of Isoelectric Focusing of Immunoglobulins.- III. Isoelectric Focusing of M-Proteins.- IV. Studies of Antibody Structure and Heterogeneity by Isoelectric Focusing.- V. Studies of Antibody Synthesis.- VI. References.- 6 The Use of Isoelectric Focusing in the Separation and Characterization of Nuclear Nonhistone Proteins.- I. Introduction.- II. General Properties of NHP.- III. Isoelectric Focusing of NHP.- IV. Artifacts and the Heterogeneity of NHP.- V. Conclusion.- VI. References.- 7 Isoelectric Focusing of Membrane Components.- I. Introduction.- II. Methods.- III. Biological Applications.- IV. References.- 8 Isoelectric Focusing of Seed Proteins.- I. Introduction.- II. Food Proteins.- III. Enzymes.- IV. Protease Inhibitors.- V. Lectins.- VI. Genetic Studies.- VII. Conclusion.- VIII. References.- 9 Isoelectric Focusing of Microbial Proteins.- I. Introduction.- II. Materials.- III. Methods.- IV. Isoelectric Focusing of Microbial Proteins.- V. Conclusions.- VI. References.- 10 Isoelectric Focusing of Serum Proteins.- I. Introduction.- II. Gel Isoelectric Focusing Alone.- III. Gel Electrofocusing Followed by Electrophoresis.- IV Individual Types of Protein in Serum or Plasma.- V. Clinical Applications.- VI. References.