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April 13, 2026Immunohematology0 citations

Effect of long-term frozen storage on antibodies in blood samples: immunoglobulin, tetanus toxin antibody, and ABO antibody

MMMasamichi MikameJapanese Red Cross Society, JapanHKHideaki KitazakiJapanese Red Cross Society, JapanRSRieko SuzukiJapanese Red Cross Society, Japan

Key Points

  • This research examines how well antibodies are preserved in frozen blood samples over long periods.
  • Compared fresh blood samples with frozen-stored samples after 14 years at -30°C
  • Measured immunoglobulin levels (IgM, IgG, IgA, IgE)
  • Evaluated tetanus toxin antibody and ABO antibody titers in the same donors using different methods
  • Used saline-direct agglutination and saline-IAT for ABO antibody assessment
  • No significant differences in overall immunoglobulin levels between fresh and frozen samples
  • Tetanus antibody titers showed preserved levels in frozen samples
  • Saline-direct agglutination ABO antibody titers were lower in frozen samples
  • Saline-IAT measured ABO antibody titers significantly decreased in frozen samples

Abstract

Abstract The Japanese Red Cross Blood Services archives donor blood samples from all donations for look-back surveys. These samples, approximately 5 million per year, are stored frozen at –30°C for 11 years and are subsequently discarded. Because such samples may also be valuable for antibody-related research and other purposes after the mandated storage period, we evaluated their potential applicability, with a particular focus on the preservation of anti- body levels. Fresh samples were compared with frozen-stored samples (–30°C for 14 years: 11 years of mandatory good manufacturing practice storage period + 3 years until measurement) for immunoglobulin levels (IgM, IgG, IgA, and IgE) as a general indicator of humoral immunity, tetanus toxin antibody titers as a long-lived vaccine-induced antibody, and ABO antibody titers (saline-direct agglutination test and saline-indirect antiglobulin test saline-IAT), which are factors in blood group typing and hemolytic transfusion reactions. For immunoglobulin levels, comparisons of non-paired fresh and frozen-stored samples were made from a random donor population (IgM, IgG, IgA: 105 cases each, IgE: 32 cases). For tetanus toxin antibody and ABO antibody, comparisons were conducted from the same donors with fresh and frozen-stored samples (tetanus toxin antibody: 47 cases, ABO antibody: 12 cases). No significant differences were observed in immunoglobulin levels, tetanus toxin antibody titers, or ABO antibody titers measured by the saline-direct agglutination method. Saline-IAT–measured ABO antibody titers were significantly lower in frozen-stored samples. Detailed results were as follows: (1) Median immunoglobulin levels (fresh vs. frozen-stored): IgM, 78.5 mg/dL versus 86.7 mg/dL; IgG, 1240.2 mg/dL versus 1280.2 mg/dL; IgA, 241.0 mg/dL versus 245.3 mg/dL; and IgE, 0.24 μg/mL versus 0.12 μg/mL. (2) Tetanus toxin antibody titers—geometric mean titer (GMT) (95% confidence interval CI): fresh 0.23 (0.12–0.43) IU/mL versus frozen-stored 0.28 (0.17–0.46) IU/mL. (3) ABO antibody titers—GMT (95% CI): saline-direct agglutination, fresh 65.4 (33.3–128.5) versus frozen-stored 51.7 (24.9–107.4); saline-IAT, fresh 391.0 (160.3–953.6) versus frozen-stored 248.7 (107.8–573.6). The evaluated immunoglobulin levels were largely preserved, suggesting that archived blood samples retain sufficient stability for antibody research. However, careful study design is necessary to minimize potential sample deterioration.

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Cite This Study

Mikame et al. (2026) studied this question.

synapsesocial.com/papers/69dc88583afacbeac03ea443https://doi.org/10.2478/immunohematology-2026-003
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