生物化学技术

Molecularly imprinted polymers (MIPs) are becoming increasingly useful as chromatographic adsorbents for molecular separations, especially chiral separations, because they can be tailored to specifically recognize the target molecule including its stereochemist ...

Chiral ligand-exchange chromatography (CLEC) first described in the late 1960s to early 1970s by Davankov and Rogozhin can be still considered as an elective choice for the direct enantioseparation of compounds endowed with chelating moieties. Among the numerous chelating species ...

The majority of enantiomeric separations for purity analysis and quality control continue to be performed by normal-phase liquid chromatography and supercritical fluid chromatography. In this chapter, representative chromatographic screening procedures for the enan ...

The risk for celiac disease (CD) is clearly related to specific HLA DQA1 and DQB1 alleles, but HLA �typing is often considered too costly for frequent use. Here we present a method using sequence-specific primed PCR (PCR-SSP) for HLA-DR-DQ genotyping optimized for capillary electrophoresis on ...

Microsatellites are short repetitive sequences of two, three, or four bases, prone to base mispairing. Microsatellite instability (MSI) occurs frequently in various types of cancer due to a defective DNA mismatch repair system. Therefore, MSI analysis is an important tool in clinical re ...

Capillary electrophoresis is an analytical technique that separates ions based on their electrophoresis mobility with the use of an applied voltage. Capillary electrophoresis is used most predominantly in nuclear acid fragment analysis as well as DNA sequencing because it gives f ...

Immunoaffinity capillary electrophoresis (ICE) is a powerful tool used to detect and quantify target proteins of interest in complex biological fluids. The target analyte is captured and bound to antibodies immobilized onto the wall of a capillary, labeled in situ with a fluorescent dye, e ...

Serum protein electrophoresis is widely used in clinical laboratories to measure the relative abundance of each obtained fraction. Moreover, we found that the migration time of the γ-globulin fraction can be reproducibly determined (CV = 1.1%). Immunoglobulins were purified from ser ...

Microfluidic immunoassay techniques offer advantages in speed, automation, and portability over �bench-top gold standard counterparts. In particular, on-chip immunosubtraction is a rapid homogeneous immunoassay used for reporting both protein native mobility and bind ...

Variations in the amino acid sequence, glycosylation, and/or other posttranslational modifications in glycoproteins give rise to different molecules of the glycoprotein called forms. Qualitative and/or quantitative alterations in these forms are related to pathophysio ...

Urine is an excellent sample source in the proteomic study of diseases. It is available in large quantities, is relatively stable, is not contaminated by cells or lipids, and has shown to provide information not only on the organs in contact with the urinary tract but also of more remote organs and tissue ...

The integration of affinity columns in microfluidic devices generates a micro-total analysis system which has high value in applications such as analyte extraction and preconcentration. In this chapter we describe the preparation of affinity columns in situ by photopolymerizat ...

Besides proteome complexity, the greatest bioanalytical challenge facing comprehensive proteomic analysis, particularly in the identification of low abundance proteins, is related to the large variation of protein relative abundances. In contrast to universally enrich ...

Protein profiling is an increasingly valuable tool for the characterization of protein populations and has been used to identify microorganisms, most often using two-dimensional gel electrophoresis followed by mass spectrometry. We present a rapid method for the identificati ...

Among the molecular diagnostic methods for bacteria-induced diseases, capillary electrophoresis-based single-strand conformation polymorphism (CE-SSCP) combined with 16S rRNA gene-specific PCR has enormous potential because it can separate sequence variants usi ...

Precise, fast, and cheap identification of pathogenic bacteria is a key factor of microbial diagnostics. Application of analytical techniques can provide new possibilities in this regard. The bacterial pathogen Staphylococcus aureus is responsible for a number of serious human in ...

Quantitative defects in hemoglobin (Hb) are represented by Hb variants, where the amino acids sequence is modified as a consequence of a mutation in the α or β-globin genes. More than 1,100 variants have been described so far but only a few dozen are clinically significant; the most significant being Hb ...

This chapter reports the essential elements needed to understand basic laboratory diagnostics consisting of separation and measurement of the hemoglobin fractions. Although well established, basic diagnostics require some background and some degree of experience for appl ...

Single-strand conformation polymorphism by capillary electrophoresis (SSCP-CE) has been developed to detect single nucleotide mutations. This method is used to identify the ABO alleles A1, A1v, B, O1, O1v, and O2 in this chapter. Four amplicons (112, 121, 123, and 160 bp) labeled with fluoresce ...

Glycosylation research has gained significant attention in several research fields including immunology, protein production, and biomarker discovery. However, complex and time-consuming protocols are often necessary to obtain suitable samples for analysis. We here desc ...