최근 4년간 수혈대상환자에서 Ortho BioVue System을 이용한 비예기항체의 빈도와 분포

임가영,박경선,박태성,이희주,서진태,박수연 대한수혈학회 2009 大韓輸血學會誌 Vol.20 No.1

배경: 최근 용혈성수혈부작용의 중요한 원인으로 비예기항체가 부각되고 있다. 비예기항체를 증명하기 위한 검사법으로 예전에는 시험관법을 많이 이용하여 왔는데 최근에는 방법이 간편하고 온난 항체의 검출율이 높은 원주응집법을 많이 이용한다. 이에 저자들은 수혈의뢰 환자들을 대상으로 최근 4년간 원주응집법으로 동정된 비예기항체의 빈도 및 분포를 알아보고 항체가 동정된 환자들의 수혈의 임상적 특성을 알아보고자 하였다. 방법: 2005년 1월부터 2008년 12월까지 경희의료원에서 수혈이 요청된 환자 44,008명의 혈청검체를 대상으로 하였다. Ortho BioVue System (Ortho-Clinical Diagnostics, NJ, USA)을 이용하여 비예기항체 선별 및 동정검사를 시행하였다. 결과: 선별검사가 의뢰된 혈청 44,008명 중 589명(1.3%)의 환자에서 비예기항체 양성반응을 보였다. 이중 383예에서 비예기항체가 동정되었는데, 가장 많은 빈도로 검출된 것은 Lewis계열, 130예(34.0%)이나 온난항체인 Rh계열과 Kidd계열이 각각 67예(17.5%), 2예(0.5%)로 동정되었다. 또한 정확하게 동정되지 않은 비예기항체는 133예(38.9%)였다. 비예기항체 선별검사에서 양성이 나와 동정된 383명의 환자의 과거력을 조사한 결과 137예(35.8%)에서 수혈기왕력이 있었고 임신이나 수혈기왕력이 있는 환자는 244예(63.7%)이었다. 결론: 한냉항체인 Lewis계열 항체가 가장 많이 동정되었고 임상적으로 의의가 있는 Rh계열의 온난 항체가 많이 검출되었다. Background: Unexpected antibodies are important factors for hemolytic transfusion reactions. In the past, the tube method was used for detecting unexpected antibodies. The column agglutination method has recently been widely used because of its simplicity and it has a higher rate of detecting warm antibodies. In this study, we describe the frequency and distribution of unexpected antibodies in transfusion candidates during the recent 4 years and the transfusion characteristics in the identified cases. Methods: Antibody screening tests were carried out on 44,008 sera using the column agglutination method from January, 2005 to December, 2008. The antibodies were screened and identified by the Ortho BioVue System (Ortho-Clinical Diagnostics, Raritan, NJ, USA). Results: Of the 44,008 cases that underwent unexpected antibodies screening, 589 cases (1.3%) showed positive results. Unexpected antibodies were identified in 383 cases. The antibodies that were most frequently detected were anti-Lewis antibodies in 130 cases (34.0%). Among the warm antibodies, anti-Rh and anti-Kidd antibodies were detected in 67 cases (17.5%) and 2 cases (0.5%), respectively. Unidentified antibodies were detected in 133 cases (38.9%). Among the patients with unexpected antibodies, 137 cases (35.8%) had a history of previous transfusion and 244 cases (63.7%) had a history of previous transfusion or gestation. Conclusion: Anti-Lewis cold antibodies were the most frequently detected antibodies. Warm antibodies were also frequently detected, and these are clinically significant.

HLA Class II 항체 검출 시 C3d 검사법과 SAB 검사법의 상관관계 평가

강혜인,김도훈,하정숙,류남희,전동석,이원목 대한진단검사의학회 2020 Laboratory Medicine Online Vol.10 No.4

Background: Detection of anti-human leukocyte antigen (HLA) antibodies is important during the selection of an appropriate donor prior to organ transplantation and also for monitoring the patients after transplantation. In this study, we compared antibodies detected via C3d assays, which monitors C3d complement-binding activities of HLA antibodies with those detected via single antigen bead (SAB) assays. Methods: A total of 66 serum samples were tested in parallel by SAB assays (Immucor Transplant Diagnostics, USA) and C3d assays (Immucor) for the detection of HLA class II antibodies. The relationship between these two methods was analyzed based on the types, numbers, median fluorescent intensity (MFI) values, and positivity of the antibodies using MATCH IT! Antibody (Immucor) program. Results: The number of antibodies obtained based on SAB and C3d assays was the highest with 24 samples (36.4%) in the 11–20 range and 23 (34.8%) in the 2–5 range detected via each assay. Among the SAB-positive antibodies, only 28 (6.4%) of the 440 antibodies with MFI ≤3,000 were C3d-positive, and 341 (61.3%) of the 556 antibodies with MFI ≥3,001 were C3d-positive. Whereas, among the 442 C3d-positive antibodies, SAB assays were positive except for 32 (7.2%) and 41 (9.3%) antibodies in the sections of MFI ≤500 and 1,001 ≤MFI ≤10,000, respectively. C3d-positive samples had higher maximum MFI values based on SAB assays, compared with C3d-negative samples. Conclusions: MFI values of HLA class II antibodies detected through SAB assays in C3d-positive samples were higher than those in C3d-negative samples. 배경: 장기이식 전에 공여자 선택을 위해 사전에 공여자 HLA에 대한 항체 유무를 확인하는 것이 중요하고, 이식 후의 급성 또는 만성 거부반응을 예측하기 위해서도 HLA 항체에 대한 주기적인 추적 검사가 필요하다. 본 연구에서는 보체 활성화를 유발하는 HLA 항체를 검출할 수 있는 Luminex 검사법 중 C3d 검사를 시행하여 기존의 SAB 검사법과 비교하였다. 방법: HLA 항체 검사를 위해 검사실로 의뢰된 검체 중 class II SAB 검사(Immucor Transplant Diagnostics, USA)에서 양성인 혈액 검체 중 최종적으로 DSA 양성인 66개의 검체를 대상으로 하였다. 각 검체를 SAB 검사법과 C3d 검사법(Immucor Transplant Diagnostics)으로 검사를 시행한 후 MATCH IT! Antibody v1.3.1.5 (Immucor Transplant Diagnostics) 프로그램으로 HLA 항체 종류, MFI 값, 양성여부를 파악한 후 두 검사의 상관관계를 분석하였다. 결과: SAB 검사의 항체수는 11-20개인 구간에서 24개(36.4%)로 가장 많았고, C3d 검사는 항체수가 2-5개인 구간에서 23개(34.8%)로 가장 많았다. SAB 검사에서 양성인 항체 중에서 MFI≤3,000인 항체 440개 중 28개(6.4%)만이 C3d 검사 양성이었고, MFI≥3,001인 항체 556개 중 341개(61.3%)가 C3d 검사 양성이었다. 반면, C3d 검사에서 양성인 442개의 항체 중에서는 MFI≤500, 1,001≤MFI≤10,000인 구간에서 각각 32개(7.2%), 41개(9.3%)를 제외하고는 모두 SAB 검사도 양성이었다. C3d 검사 양성 검체가 C3d 검사 음성 검체보다 SAB의 최대 MFI 값이 더 큰 것으로 나타났다. 결론: HLA class II 항체의 SAB 검사에서의 MFI 값은 C3d 검사 음성인 경우보다 양성인 경우에 더 큰 것으로 나타났으며, C3d 검사법의 임상적 중요성에 대해서는 추가적인 연구가 필요할 것이다.

Application of Single-Domain Antibodies (“Nanobodies”) to Laboratory Diagnosis

Pillay Tahir S.,Muyldermans Serge 대한진단검사의학회 2021 Annals of Laboratory Medicine Vol.41 No.6

Antibodies have proven to be central in the development of diagnostic methods over decades, moving from polyclonal antibodies to the milestone development of monoclonal antibodies. Although monoclonal antibodies play a valuable role in diagnosis, their production is technically demanding and can be expensive. The large size of monoclonal antibodies (150 kDa) makes their re-engineering using recombinant methods a challenge. Single-domain antibodies, such as “nanobodies,” are a relatively new class of diagnostic probes that originated serendipitously during the assay of camel serum. The immune system of the camelid family (camels, llamas, and alpacas) has evolved uniquely to produce heavy-chain antibodies that contain a single monomeric variable antibody domain in a smaller functional unit of 12–15 kDa. Interestingly, the same biological phenomenon is observed in sharks. Since a single-domain antibody molecule is smaller than a conventional mammalian antibody, recombinant engineering and protein expression in vitro using bacterial production systems are much simpler. The entire gene encoding such an antibody can be cloned and expressed in vitro. Single-domain antibodies are very stable and heat-resistant, and hence do not require cold storage, especially when incorporated into a diagnostic kit. Their simple genetic structure allows easy re-engineering of the protein to introduce new antigen-binding characteristics or attach labels. Here, we review the applications of single-domain antibodies in laboratory diagnosis and discuss the future potential in this area.

One target, different effects: a comparison of distinct therapeutic antibodies against the same targets

Shim, Hyun-Bo Korean Society for Biochemistry and Molecular Bion 2011 Experimental and molecular medicine Vol.43 No.10

To date, more than 30 antibodies have been approved worldwide for therapeutic use. While the monoclonal antibody market is rapidly growing, the clinical use of therapeutic antibodies is mostly limited to treatment of cancers and immunological disorders. Moreover, antibodies against only five targets (TNF-${\alpha}$, HER2, CD20, EGFR, and VEGF) account for more than 80 percent of the worldwide market of therapeutic antibodies. The shortage of novel, clinically proven targets has resulted in the development of many distinct therapeutic antibodies against a small number of proven targets, based on the premise that different antibody molecules against the same target antigen have distinct biological and clinical effects from one another. For example, four antibodies against TNF-${\alpha}$ have been approved by the FDA - infliximab, adalimumab, golimumab, and certolizumab pegol - with many more in clinical and preclinical development. The situation is similar for HER2, CD20, EGFR, and VEGF, each having one or more approved antibodies and many more under development. This review discusses the different binding characteristics, mechanisms of action, and biological and clinical activities of multiple monoclonal antibodies against TNF-${\alpha}$, HER-2, CD20, and EGFR and provides insights into the development of therapeutic antibodies.

적혈구 불규칙항체 션별검사의 평가

오현숙 ( Hyun Sook Oh ),최영미 ( Young Mi Choi ),김희순 ( Hee Soon Kin ),양경섭 ( Kyung Sub Yang ),김재우 ( Jea Woo Kim ) 대한임상검사과학회 1998 대한임상검사과학회지(KJCLS) Vol.30 No.3

The objective of this study was to detection irregular antibodies among 53 sera by the use of the conventional tube test, low ionic strength solution anti-human globulin(LISS-AHG) test, polyethylene glycol anti-humanglobulin(PEG-AHG) test, gel lowionic strength solution(GEL-LISS) test, and gel papain(Gel-PAP) test. these tests were used to compare their sensitivities and reactivities for the detections. The sera were collected from three hospitals and two blood centers and all were identified to have alloantibodies. Out of 27 clinically significant cases, the Gel-PAP test was 100% sensitive in detecting Rh antibodies, the PEG-AHG test was sensitive in detecting Duffy, Kidd, Ss antibodies but not Rh antibodies, and the Gel-LISS test was determined to be sensitive for Rh, Duffy, Kidd, Ss antibodies. The reactivities of these tests for the detection of clinically significant antibodies were well correlated with those of sensitivities. The other 26 sera contained clinically insignificant antibodies (anti-Le", -Leb, -M, -PI) and these were detected completely by the tube test. In our experience, Gel-LISS was more sensitive than other antibody screening tests indetecting clinically significant antibodies. but we recommend combining. methods, for example either the PEG-AHG and Enxyme method together, or the Gel-LISS and the Gel-PAP for more reliable the detection of clinically significant antibodies in sera.

One target, different effects: a comparison of distinct therapeutic antibodies against the same targets

심현보 생화학분자생물학회 2011 Experimental and molecular medicine Vol.43 No.10

To date, more than 30 antibodies have been approved worldwide for therapeutic use. While the monoclonal antibody market is rapidly growing, the clinical use of therapeutic antibodies is mostly limited to treatment of cancers and immunological disorders. Moreover, antibodies against only five targets (TNF-α, HER2, CD20,EGFR, and VEGF) account for more than 80 percent of the worldwide market of therapeutic antibodies. The shortage of novel, clinically proven targets has resulted in the development of many distinct therapeutic antibodies against a small number of proven targets,based on the premise that different antibody molecules against the same target antigen have distinct biological and clinical effects from one another. For example,four antibodies against TNF-α have been approved by the FDA -- infliximab, adalimumab, golimumab, and certolizumab pegol -- with many more in clinical and preclinical development. The situation is similar for HER2, CD20, EGFR, and VEGF, each having one or more approved antibodies and many more under development. This review discusses the different binding characteristics, mechanisms of action, and biological and clinical activities of multiple monoclonal antibodies against TNF-α, HER-2, CD20, and EGFR and provides insights into the development of therapeutic antibodies.

Rh계 항체 검출을 위한 여러 가지 검사법의 평가

오현숙 ( Hyun Sook Oh ),이명재 ( Myung Jae Lee ),이미자 ( Mi Ja Lee ),이영진 ( Young Jin Lee ) 대한임상검사과학회 2000 대한임상검사과학회지(KJCLS) Vol.32 No.3

Rh system antibodies are frequently implicated in hemolytic disease of the newbom and acute or delayed hemolytic transfusion reaction. To prevent these diseases, irregular antibodies must be screened and identified. Detection of Rh system antibodies is usually done by conventional serological methods. Recently, flow cytomeσy has been introduced in immunohemat이ogy to detect antigens and antibodies. We evaluated the possibility of clinical usage to detect Rh system antibodies by flow cytometry. We examined 6 specimens of Rh antibodies (1 anti-C+e, 2 anti-D, 2 anti-E, 1 anti-E+c) that were quantified of Rh antibodies by conventional serological methoru잉 (antiglobulin test, PEG test, gel card test, papain treated gel card test) and flow cytometer, and compared the results. The papain treated gel card test showed the highest sensitivity method to detect Rh antibodies. The resu1t of flow cytometer was well correlated with conventional serological methods especially gel card test, except in 3 samples (shown high titers). The results of flow cytometer were objective and reliable for detection and quantification of Rh antibodies. Discrepancies in the results of this study suggest that seem to be influenced by antibodies of high concentration in serum samples.

Antibody Engineering for the Development of Therapeutic Antibodies

Hyo Jeong Hong,Youngwoo Park,Sang Jick Kim 한국분자세포생물학회 2005 Molecules and cells Vol.20 No.1

Therapeutic antibodies represent one of the fastest growing areas of the pharmaceutical industry.There are currently 19 monoclonal antibodies in the market that have been approved by the FDA and over 150 in clinical developments. Driven by innovation and technological developments, therapeutic antibodies are the second largest biopharmaceutical product category after vaccines. Antibodies have been engineered by a variety of methods to suit a particular therapeutic use. This review describes the structural and functional characteristics of antibody and the antibody engineering for the generation and optimization of therapeutic antibodies.

Anti-idiotypic Antibodies against Bovine Growth Hormone

Verma, N.K.,Sodhi, R.,Rajput, Y.S. Asian Australasian Association of Animal Productio 2003 Animal Bioscience Vol.16 No.5

Anti-antibodies against three mouse monoclonal antibodies viz. IIB5D6, VIA6E8 and VIC1F9 (specific to bovine growth hormone) in rabbits have been generated and characterized. Ammonium sulfate fractionated and affinity-purified monoclonal antibodies were used for producing anti-antibodies. The generated anti-antibodies were against common as well as uncommon antigenic determinants present in mouse monoclonal antibodies. The raised anti-antibodies replaced [$I^125$ ]bGH bound to goat liver microsomes indicating production of anti-idiotypic antibodies against bovine growth hormone. These antibodies can have profound implications in vivo in lactating bovines for enhancing milk yield.

Frequency of unexpected antibody and consideration during transfusion

Ki-Ho Ko,Byung Hoon Yoo,김계민,Woo-Yong Lee,연준흠,Ki-Hyuk Hong,한태희 대한마취통증의학회 2012 Korean Journal of Anesthesiology Vol.62 No.5

Background: In this retrospective study, we measured the frequency of unexpected antibodies in the blood. Specific considerations for preoperative preparations were kept in mind for the patients undergoing surgery positive for these antibodies. Methods: After reviewing the results of antibody screening tests lasted for 2 years, the frequency of unexpected antibodies was determined. Surgical patients who were positive for unexpected antibodies were selected and divided into two groups based on their potential need for an intra-operative transfusion (groups with high versus low possibility of transfusion). Blood for the high possibility group was prepared before surgery. For the low possibility group for which preoperative blood preparation was not performed, cases of this group were reviewed whether a blood preparation was delayed or not in case of transfusion. Results: Among a total 22,463 cases, 340 (1.52%) had positive results for antibody screening tests. Among the 243 patients who were positive for unexpected antibodies, Lewis, Rh, Xga, and mixed antibodies were found in 85, 25, five, and eight cases, respectively. Out of 243 patients, 117 patients, specificities of the unexpected antibodies were not determined and 125 (51.4%) had a history of pregnancy and delivery, and 49 (20.2%) had a history of transfusion. In the low probability group, transfusions were administered for nine patients; transfusion was delayed for two patients due to difficulties with obtaining matched blood. Conclusions: Patients with unexpected blood antibodies may be at increased risk for delayed transfusion. For rapid transfusion, it might be helpful to keep a record about blood antibodies and introduce a notification system such as medical alert cards. Preoperative blood preparation is needed for timely intraoperative transfusion.