Thesis

Optimization of the Protocol for Human Immunoglobulin G (IgG) Extraction from Dried Blood Spots

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Existing protocols for extracting Immunoglobulin G (IgG) from blood spots (DBS) were compared,
utilizing fresh, one-week-old, and one-month-old DBS made from a healthy Thai’s blood as the
samples. In addition, DBS samples from healthy uninfected villagers obtained from a Malaria survey in
2017 will also be used. To validate the previously published elution protocols and buffers, alterations
of elution buffers were applied in a down-selection approach, uncovering the optimal buffer
compositions that can elute the highest yield and purity of IgG. The IgG level in the eluates obtained
from the elution using NP-40, particularly 0.5% NP-40, which showed the strongest activity in IgG
elution, was significantly distinct with the IgG level obtained using the 0.05% Tween-20 buffer, which
is the current standard protocol applied in Mahidol Vivax Research Unit (MVRU). While all protocols
can relatively extract the IgG from DBS samples, only buffers that contain either NP-40 or ammonium
chloride, aside from PBS, can successfully extract IgG from seven-years-old DBS samples at a
significant level. In addition, all of the tested protocols were done in the most effective manner,
requiring only a low cost, simple equipment, and a short time to be performed in order to find the
most applicable protocol that can be utilized during emergency situations in rural areas.

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Availability

Detail Information

Series Title

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Call Number

BM 24-039

Publisher

i3L, Jakarta : i3L Press., 2024

Collation

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Language

English

ISBN/ISSN

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Classification

NONE

Content Type

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Media Type

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Carrier Type

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Edition

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Subject(s)

ELISA
dried blood spots (DBS)
elution
immunoglobulin
IgG

Specific Detail Info

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Statement of Responsibility

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Other version/related

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