"Lyophilization of Immunocomponents: A Technical Approach for Preservation and Application in Equine Therapies."
Immunoglobulins; Platelets; Spray dryer; Hyperimmune plasma; Equine plasma.
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The use of lyophilized compounds represents a practical and accessible solution for the transport and storage of immunocompounds in equine medicine. This thesis aims to evaluate the technical viability of the lyophilization and atomization process of blood plasma as a source of immunocomponents. In the first chapter, we measured the concentrations of immunoglobulin G (IgG), total protein (TP), and total solids (TS) in fresh equine plasma and after lyophilization. Plasma was collected from six healthy male horses, lyophilized, and reconstituted in deionized water to its original volume. The IgG concentrations in fresh plasma (8.9 ± 3.2 g/L) and lyophilized plasma (7.1 ± 2.2 g/L) showed no significant difference (P > 0.05). In contrast, the TP concentration decreased from 6.6 ± 0.5 g/dL in fresh plasma to 5.7 ± 0.2 g/dL after lyophilization (P < 0.05), and the TS was also reduced from 7.5 ± 0.8% to 6.3 ± 0.5% (P < 0.05). These results indicate that lyophilization preserves the IgG concentration, with minor losses in TS and TP. The second chapter evaluated the integrity of platelets in lyophilized, spray-dried, and frozen equine plasma compared to fresh plasma. Blood samples were collected from six healthy male horses in tubes containing sodium citrate, and the obtained plasma underwent freezing at -80°C, lyophilization, and spray-drying. Platelet integrity was assessed before and after reconstitution using a hematology analyzer, and the concentrations of LDH, TS, and PT were measured. The average platelet count in fresh plasma was 125 ± 49.90 x 10^9/L, with lyophilization resulting in a 46.16% reduction (67.3 ± 30.72 x 10^9/L; P < 0.05). Despite this decrease, 53.84% of the platelets remained intact. Freezing led to a 30.64% reduction in count (86.7 ± 32.85 x 10^9/L; P < 0.05). The concentrations of LDH, PT, and TS did not differ significantly between the groups. In contrast, spray-drying caused irreparable damage to the platelets, resulting in a count of zero after reconstitution. This study illustrates the impact of lyophilization, spray-drying, and freezing on equine plasma, particularly regarding platelet count, total protein, and LDH levels. While lyophilization and freezing preserved platelet integrity to some extent, spray-drying was ineffective. The findings highlight the need to optimize preservation methods to enhance the stability and therapeutic potential of equine plasma in veterinary medicine. The research supports the use of lyophilized equine plasma as a promising treatment option, with future investigations focused on validating its efficacy and stability, as well as developing practical packaging solutions for the equine industry.
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