Determination of lectin characteristics by a novel agglutination technique

Flemming, C; Ulrich, Michael; Schwall, Gerhard P.; Schröder, Heinz C.; Müller, Wernér E.G.

doi:10.1016/0003-2697(92)90431-6

Cited by 5 publications

(2 citation statements)

References 14 publications

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“…Flemming et al designed a simple apparatus called agglutimeter which utilized a photometer for measurement of turbidity of samples containing agglutinates. This technique can be applied not only to agglutination of erythrocytes but all types of cells [49]. Optical density, turbidity or dynamic light scattering were measured when agglutination assays with glycan-decorated vesicles was used for investigation of lectin multivalent interactions [50] or lectin hetero-multivalency [51].…”

Section: Resultsmentioning

confidence: 99%

Microscopy examination of red blood and yeast cell agglutination induced by bacterial lectins

2019

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Lectins are a group of ubiquitous proteins which specifically recognize and reversibly bind sugar moieties of glycoprotein and glycolipid constituents on cell surfaces. The mutagenesis approach is often employed to characterize lectin binding properties. As lectins are not enzymes, it is not easy to perform a rapid specificity screening of mutants using chromogenic substrates. It is necessary to use different binding assays such as isothermal titration calorimetry (ITC), surface plasmon resonance (SPR), microscale thermophoresis (MST), enzyme-linked lectin assays (ELLA), or glycan arrays for their characterization. These methods often require fluorescently labeled proteins (MST), highly purified proteins (SPR) or high protein concentrations (ITC). Mutant proteins may often exhibit problematic behaviour, such as poor solubility or low stability. Lectin-based cell agglutination is a simple and low-cost technique which can overcome most of these problems. In this work, a modified method of the agglutination of human erythrocytes and yeast cells with microscopy detection was successfully used for a specificity study of the newly prepared mutant lectin RS-IIL_A22S, which experimentally completed studies on sugar preferences of lectins in the PA-IIL family. Results showed that the sensitivity of this method is comparable with ITC, is able to determine subtle differences in lectin specificity, and works directly in cell lysates. The agglutination method with microscopy detection was validated by comparison of the results with results obtained by agglutination assay in standard 96-well microtiter plate format. In contrast to this assay, the microscopic method can clearly distinguish between hemagglutination and hemolysis. Therefore, this method is suitable for examination of lectins with known hemolytic activity as well as mutant or uncharacterized lectins, which could damage red blood cells. This is due to the experimental arrangement, which includes very short sample incubation time in combination with microscopic detection of agglutinates, that are easily observed by a small portable microscope.

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Section: Resultsmentioning

confidence: 99%

Microscopy examination of red blood and yeast cell agglutination induced by bacterial lectins

2019

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“…In this way, different strains of non‐pathogenic yeasts of the same species can be distinguished rapidly and exactly as Flemming et al . stated in their examination of Saccharomyces cerevisiae isolates by investigation of strain‐specific lectin‐mediated (LM) agglutination kinetics [20–23].…”

Section: Introductionmentioning

confidence: 99%

Differentiation of Candida strains by lectin‐mediated agglutination kinetics

Nenoff

Süss

Flemming

et al. 2000

Mycoses

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The lectin-mediated agglutination kinetics of Candida albicans, Candida tropicalis, Candida glabrata, Candida krusei, Candida kefyr, and Candida parapsilosis strains isolated from immunocompromised patients was investigated. The rate of the lectin-induced cell agglutination depends on the physiological state of the yeast cell population. Therefore, the Candida strains have to be cultivated and investigated under identical conditions. Lentil lectin (prepared from Lens culinaris), castor lectin, and concanavalin A were used. Different yeast species showed different agglutination behaviour. Furthermore, the lectin-mediated rate of agglutination is a strain-specific property which makes it possible to distinguish between different yeast strains of the same species. It is concluded that the lectin-mediated agglutination kinetics allows reproducible differentiation of yeast strains of the same species.

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Quantitative Estimation of Coproporphyrin III and Protoporphyrin IX from Their Mixture by Spectrofluorometry

Tewari

Tripathy

1998

J. Plant Biochem. Biotechnol.

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Determination of lectin characteristics by a novel agglutination technique

Cited by 5 publications

References 14 publications

Microscopy examination of red blood and yeast cell agglutination induced by bacterial lectins

Microscopy examination of red blood and yeast cell agglutination induced by bacterial lectins

Differentiation of Candida strains by lectin‐mediated agglutination kinetics

Quantitative Estimation of Coproporphyrin III and Protoporphyrin IX from Their Mixture by Spectrofluorometry

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