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ASTM E1470-92(1998)

(Test Method)

Standard Test Method for Characterization of Proteins by Electrophoretic Mobility

Standard Test Method for Characterization of Proteins by Electrophoretic Mobility

SCOPE
1.1 This test method describes a procedure for determining the electrophoretic mobility of proteins of molecular weight greater than 10000 Daltons.  
1.2 This test method uses automatic Electrophoretic Light Scattering (ELS) principles to determine the electrophoretic mobility.  
1.3 The instrument  simultaneously measures the Doppler shifts of scattered light at four different angles to determine the electrophoretic mobility distribution of protein particles. The mobility is expressed as [mu]m-cm/V-s (micron-centimeter/volt-second).  
1.4 This standard does not purport to address all of the safety problems, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

General Information

Status
Historical
Publication Date
09-Apr-1998
ICS
07.080 - Biology. Botany. Zoology
Technical Committee
E48 - Bioenergy and Industrial Chemicals from Biomass
Current Stage
Ref Project

Relations

Replaced By
ASTM E1470-92(2006) - Standard Test Method for Characterization of Proteins by Electrophoretic Mobility (Withdrawn 2014)
Effective Date
01-Nov-2006
Referred By
ASTM F594-22 - Standard Specification for Stainless Steel Nuts
Effective Date
10-Apr-1998
Referred By
ASTM E2865-12(2022) - Standard Guide for Measurement of Electrophoretic Mobility and Zeta Potential of Nanosized Biological Materials
Effective Date
10-Apr-1998

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ASTM E1470-92(1998) - Standard Test Method for Characterization of Proteins by Electrophoretic MobilityASTM E1470-92(1998) - Standard Test Method for Characterization of Proteins by Electrophoretic Mobility
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ASTM E1470-92(1998) - Standard Test Method for Characterization of Proteins by Electrophoretic Mobility
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NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
Designation:E1470–92(Reapproved 1998)
Standard Test Method for
Characterization of Proteins by Electrophoretic Mobility
This standard is issued under the fixed designation E 1470; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (e) indicates an editorial change since the last revision or reapproval.
1. Scope 3.2 Both sellers and purchasers of protein particles will find
this test method useful to determine either mobility or zeta
1.1 This test method describes a procedure for determining
potentialdistributionsforproteinspecifications,manufacturing
the electrophoretic mobility of proteins of molecular weight
control, and development and research.
greater than 10 000 Daltons.
1.2 This test method uses automatic Electrophoretic Light
4. Apparatus
Scattering (ELS) principles to determine the electrophoretic
4.1 The apparatus for analysis consists essentially of a laser
mobility.
2 light source, sample cell for introducing the sample, power
1.3 The instrument simultaneously measures the Doppler
supply source, four 256 channel spectrum analyzers, micropro-
shifts of scattered light at four different angles to determine the
cessors, and computer assembly.
electrophoretic mobility distribution of protein particles. The
4.2 Sample chamber assembly, holds approximately 1 mL
mobility is expressed as µm-cm/V-s (micron-centimeter/volt-
of sample and is composed of three basic parts. The two side
second).
pieces are made of solid silver and contain hemispherical
1.4 This standard does not purport to address all of the
cavities. Between the two side pieces is a fused silica glass
safety concerns, if any, associated with its use. It is the
insert, running through it is a rectangular channel (3 mm wide
responsibility of the user of this standard to establish appro-
by 1 mm high). The channel connects the two cavities. Fluid
priate safety and health practices and determine the applica-
fills both cavities and the channel. Electrophoretic Light
bility of regulatory limitations prior to use.
Scattering measurements are made on particles in the channel.
2. Summary of Test Method 4.2.1 30mLPlasticAccuvetts,(disposable)forpreparingthe
sample.
2.1 A carefully dispersed, dilute suspension of the protein
4.2.2 Membrane Filtering Device, 0.2 µm filters or finer.
particles is loaded into the sample cell and is positioned in the
4.2.3 5 mL Sterile Plastic Syringe.
path of collimated laser light. The laser light directed onto
4.2.4 8 Gage Blunt Tipped Hypodermic Needle.
particles moving at constant velocity under an applied electri-
4.2.5 pH Meter.
cal field. The laser light is scattered from moving particles,
4.2.6 Standard Buffer Solution.
producing a Doppler shift proportional to the particle’s veloc-
ity.
5. Reagents and Materials
2.2 The instrument response is essentially to a sinusoidal“
5.1 Purity of Reagents—Reagent grade chemicals shall be
beat” signal produced at the detector by mixing the scattered
used in all tests. Unless otherwise indicated, it is intended that
light and a reference (unscattered) beam. The frequency of the
all reagents shall conform to the specifications of the Commit-
“beat” signal is equal to the difference Doppler shift and
tee on Analytical Reagents of the American Chemical Society
therefore, to particle speed and direction.
where specifications are available. Other grades may be used,
3. Significance and Use provided it is first ascertained that the reagent is of sufficiently
high purity to permit its use without lessening the accuracy of
3.1 The prime purpose of this test method is to provide data
the determination.
expressed as either electrophoretic mobility or zeta potential
5.2 Suspending Media—The sample media could be any
distribution of protein particles.
standard buffer solution (conductivity 2 µs to 200 millisiemen).
The media shall be filtered through 0.2 µm or finer membrane
This test method is under the jurisdiction of ASTM Committee E-48 on
Biotechnology and is the direct responsibility of Subcommittee E48.03 on Unit
Processes and Their Control.
Current edition approved March 15, 1992. Published May 1992. “Reagent Chemicals,American Chemical Society Specifications,”Am. Chemi-
The Coultert Delsa 440 instrument from Coulter Corporation has been found cal Soc., Washington, DC. For suggestions on the testing of reagents not listed by
satisfactory. This instrument is available from Coulter Corporation, 601 W. Coulter the American Chemical Society, see “Analar Standards for Laboratory U.K.
Way, Hialeah, FL 33010. Chemicals,” BDH Ltd., Poole, Dorset, and the “United States Pharmacopeia.”
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
...

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