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Measuring of Alpha-1 Antitrypsin Concentration by Nephelometry or Turbidimetry

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Alpha-1 Antitrypsin

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2750))

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Abstract

Most clinical laboratories quantify alpha-1 antitrypsin using either nephelometry or turbidimetry techniques because they are commercially available, amenable to automation, and precise. Both methods are based on light scatter. The foundation of both techniques is based on incubation of the specimen with anti-AAT polyclonal antibody solution, a polymer matrix between endogenous AAT and the reagent antibodies forms, leading to production of light-scattering large particles. Although these two terms are sometimes used synonymously, technically speaking they are not.

Nephelometry measures the amount of turbidity or cloudiness of a solution by directly quantifying the intensity of the light scattered by insoluble particles in the sample. Therefore, this technique measures the light that passes through the sample, with the detector being placed at an angle from the sample. Turbidimetry is the process of measuring the loss of intensity of the light transmitted linearly through a sample caused by the scattering effect of insoluble particles. The decrease in light transmission is measured compared to a reference, and the absorbed light is quantified.

Beyond specific technical differences between both techniques, there are two major differences between the two procedures that may influence the results. First, the concentration of the sample and the resulting intensity of scattered light relative to the intensity of the light source is one major factor. Second, the size of the scattering particles is also a key differentiating factor. This chapter describes the technical requirements, the different protocols, and the clinical applicability of these two techniques in the diagnosis of alpha-1 antitrypsin deficiency.

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Authors and Affiliations

  1. Unidad Médico-Quirúrgica de Enfermedades Respiratorias, Instituto de Biomedicina de Sevilla (IBiS), Hospital Universitario Virgen del Rocío/CSIC/Universidad de Sevilla, Seville, Spain

    Carmen Marin-Hinojosa & Jose Luis Lopez-Campos

  2. CIBER de Enfermedades Respiratorias (CIBERES), Instituto de Salud Carlos III, Madrid, Spain

    Carmen Marin-Hinojosa & Jose Luis Lopez-Campos

  3. Unidad de Gestión Clínica de Laboratorios, Servicio de Bioquímica Clínica, Sección de Inmunoproteínas y Marcadores Tumorales, Hospital Universitario Virgen del Rocío/Universidad de Sevilla, Seville, Spain

    Daniel Fatela-Cantillo

Authors
  1. Carmen Marin-Hinojosa
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  2. Daniel Fatela-Cantillo
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  3. Jose Luis Lopez-Campos
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Corresponding author

Correspondence to Jose Luis Lopez-Campos .

Editor information

Editors and Affiliations

  1. Alpha-1 Biologics, Long Island High Technology Incubator, Stony Brook University, Stony Brook, NY, USA

    Cynthia L. Bristow

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Marin-Hinojosa, C., Fatela-Cantillo, D., Lopez-Campos, J.L. (2024). Measuring of Alpha-1 Antitrypsin Concentration by Nephelometry or Turbidimetry. In: Bristow, C.L. (eds) Alpha-1 Antitrypsin. Methods in Molecular Biology, vol 2750. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3605-3_12

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  • DOI: https://doi.org/10.1007/978-1-0716-3605-3_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3604-6

  • Online ISBN: 978-1-0716-3605-3

  • eBook Packages: Springer Protocols

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