We intended to reformulate an existing
platelet-derived wound healing formula to target each phase of the healing wound with the appropriate phase-specific molecules. A decreased perfusion of the skin, often associated with conditions such as
thalassemia,
sickle cell disease,
diabetes mellitus, and chronic
vascular disease, is the most common etiology of cutaneous
ulcers and chronic
wounds. We had previously shown that a
PDWHF topically applied to a chronic nonhealing
ulcer of a β-
thalassemia homozygote stimulated and accelerated closure of the
wound. The
PDWHF was prepared from a pooled platelet concentrate of a matching
blood group, consisting of a combination of platelet α-granule-derived factors. Processing of the
apheresis-pooled platelets yielded various amounts of
proteins (3.36 g/mL ± 0.25 (SD) (N = 10)) by the better lysis
buffer method.
Immunoglobulin G was found to be the most abundant α-granule-secreted
protein. Equally broad quantities of the
IgG (10.76 ± 12.66% (SD) (N = 10)) and
IgG/
albumin ratios (0.6 ± 0.4 (SD) (N = 10)) were quantified. We have developed a method using a reformulated lysis
buffer followed by size exclusion chromatography and affinity chromatography to extract, identify, quantify, and purify
IgG from activated platelets.
IgG purification was confirmed by Western blot and flow cytometry. It was thought unlikely that the platelet
IgG could be accounted for by adsorption of
plasma protein, though the variable quantities could account for diversity in wound healing rates. The
IgG could protect the
wound even from
subclinical infections and functionally advance healing. It may be useful in the management of
skin ulcers in the early phase of wound healing.