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Mahbub Hasan,Jong-Gu Choi,Hafeza Akter,Sang-Suk Lee 한국자기학회 2022 Journal of Magnetics Vol.27 No.4
The suppression of cytokine storm in severe coronavirus disease 2019 (COVID-19) patients can be treated with monoclonal antibody therapy against CD3 for T cell receptor inhibition. An optimized liquid phase as a CD3 antibody-magnetic nanoparticle (Ab-MNP) conjugate can inhibit the overactivation of T cells. We aim to analyze the distribution of Fe in the spleen after acute administration of silica-conjugated amine magnetite (Fe3O4) nanoparticles (35 nm) delivered by intravenous injection. The Fe element distribution and concentration levels in spleen tissue were analyzed using energy dispersive spectroscopy (EDS) and inductively coupled plasmamass spectrometry (ICP-MS). The experimental result is a difference in the concentration of Fe elements, which was 1.89×103 mg/kg in the spleen of a control mouse not administered with MNPs, whereas increases significantly to 1.93×103 mg/kg in that of a mouse administered with MNPs. Further, time kinetic analysis of biochemical and immunological parameters is required to confirm its suitability in bio-administration.
항-CD3 단일클론 항체와 접합된 아민-덱스트란 자성나노입자의 생리식염수 내 자기저항 특성
HASAN MAHBUB,최종구,신명우,이상석 한국물리학회 2023 새물리 Vol.73 No.9
COVID-19의 사이토카인 폭풍의 원인이 되는 T 세포 과잉 활성화를 억제하는 면역치료용 항체인 항-CD3e mAb(145-2C11)를 자성나노입자(MNPs)의 표면에 접합하여 자기적 특성을 조사하였다. MNPs와 CD3 항체의 접합에 필요한 가교제로 글루타르알데하이드를 이용하였다. TEM-EDS 스펙트럼 분석에서 존재하는 원소의 무게 순서는 C > O > Fe > P > Cl > N > S 이었으며, 2단자 구리 전극 수조에서 측정한 MNPs-mAb 접합체가 포함된 PBS 용액의 자기저항 곡선으로부터 자기저항값, 자기저항비, 용액 보자력은 각각 18 MΩ, −22.2%, 300 Oe으로 대칭 피크를 나타내었다. 이러한 결과는 항체가 접합된 MNPs가 외부 자기장에 의하여 원하는 표적으로 조절 및 유도될 수 있음을 보여주는 것으로 사료된다. Anti-CD3e mAb (145-2C11), which is an immunotherapeutic antibody that inhibits the overactivation of T cells and the cause of the cytokine storm in COVID-19, was conjugated to the surface of magnetic nanoparticles (MNPs), and its magnetic properties were investigated. Glutaraldehyde was used as a cross-linking agent to conjugate MNPs and CD3 antibody (MNP-mAb). In the transmission electron microscopy–energy dispersive spectroscopy spectrum analysis, the order of the weights of the elements present was C > O > Fe > P > Cl > N > S. From the magnetoresistance curve of the phosphate-buffered saline solution containing the MNP-mAb conjugate measured in a two-terminal copper electrode bath, the magnetoresistance value, ratio, and solution coercivity were 18 MΩ, −22.2%, and 300 Oe, respectively, with symmetrical peaks. These results reveal that the antibody-conjugated MNPs can be regulated and induced to the desired target using an external magnetic field.
용액 안에 담겨 있는 항체와 결합용 자성나노입자의 자기저항 특성
이상석,최종구,Mahbub Hasan,Hafeza Akter,이현숙 한국자기학회 2022 韓國磁氣學會誌 Vol.32 No.3
Magnetic nanoparticles with an average diameter of less than 50 nm with an amine-group surface coated with silica or dextran are being used as immunotherapeutic antibody conjugates. The magnetoresistance (MR) characteristics of magnetic nanoparticles (MNPs) are investigated according to the strength of the external magnetic field that changed the distribution of the nanoparticle solution at a 1 mm interval between two copper electrodes having a diameter of 1 mm. In the case of the silica amine group MNP solution, which is rapidly separated by an external magnetic field, the magnetoresistance curve is non-uniform. In the case of MNPs with an antibody attached to an amine group, the initial MR value, MR ratio, and solution coercivity are 0.74M, 1.6%, and 200 Oe, respectively, depending on the external magnetic field. In addition, it took more than 10 hours under a powerful magnetic field to completely separate the magnetic dextran nanoparticles from which the dispersant is sufficiently added to a MNP solution of 2 mg/mL. The dextran MNP solution, which maintained the concentration, shows a symmetric butterfly-shaped magnetoresistance curve with MR, MR ratio, and solution coercive force of 1.43M, 0.2%, and 260 Oe, respectively, depending on the external magnetic field. These results show that it is possible to control the induction of MNPs for the development of antibodies for immunotherapy to a target with a magnetic field.
자성나노입자 주입에 의한 혈청의 인터페론-감마 변화 특성
이한승,전찬호,HASAN MAHBUB,최종구,AKTERHAFEZA,배예빈,이상석 한국물리학회 2022 새물리 Vol.72 No.9
Magnetic nanoparticles are widely used for treatment and diagnosis in the medical field. Given that their position is easily manipulated using a magnetic field, they can be developed as targeted immunotherapeutic agents. This study aims to determine whether dextran coat on 40 nm Fe3O4 magnetic nanoparticles is medically toxic. The interferon-gamma (IFN-γ) concentration is measured using a sandwich ELISA method. IFN-γ is a potent inflammatory cytokine that increases with toxic immune responses. If magnetic nanoparticles are toxic, then an inflammatory reaction occurs when they are injected into the body through the tail vein of a mouse, resulting in a change in IFN-γ concentration in blood serum. However, when dextran-coated magnetic nanoparticles are injected in vivo, the IFN-γ concentration in the mouse serum does not increase, so an inflammatory reaction does not occur. Therefore, the dextran-coated magnetic nanoparticles are biocompatible and suitable for antibody-conjugated targeted immunotherapeutic drug development. 치료와 진단의 목적으로 의료분야에 많이 사용되고 있는 자성나노입자는 자기장에 의해 위치조작이 가능한 장점이 있어 표적 면역 치료제로 개발될 수 있다. 40 nm 크기의 자철석 Fe3O4 에 덱스트란이 코팅된 자성나노입자를 의학적으로 독성 여부를 확인하기 위해 샌드위치 ELISA 방법을 이용하여 인터페론-감마 (IFN-γ)의 농도를 측정하게 된다. 자성나노입자가 독성이 있다면 마우스 꼬리 정맥을 통해 체내 주입되었을 때 염증 반응이 일어나 혈액의 혈청 내 IFN-γ 농도의 변화를 나타내게 된다. 면역 반응이 일어나면 사이토카인이 분비되기 때문에 IFN-γ가 증가하게 된다. 덱스트란-자성나노입자를 생체 내에 주입하였을 때 마우스의 혈청 내의 IFN-γ가 증가하지 않았기에 염증 반응이 일어나지 않아서 독성이 없는 생체적합성 물질로 면역치료에 적합한 단일클론항체를 결합한 자성나노입자를 의약학적으로 사용이 가능할 수 있다.
유도결합 플라즈마 질량분석법을 이용한 마우스 비장 내 Fe 입자 분포 연구
임예은,이정인,HASAN MAHBUB,최종구,AKTERHAFEZA,이상석 한국물리학회 2022 새물리 Vol.72 No.9
Magnetic nanoparticles (MNPs) are biocompatible materials. One example is iron (Fe), one of 14 trace elements constituting living things. MNPs are used in various bioapplications comprising drug delivery and thermotherapy. After anesthesia, mice were injected intravenously (tail vein) with either 50 μL of PBS (control group) or MNPs (treatment group) and maintained for 20 days. The MNPs were administered at a concentration of 2 μg/μL. The mice were sacrificed on days 0, 5, 10, 15, and 20 post-MNP injections. After anatomical examination, the organs were collected and wet weights were measured. The spleen, which stores immune cells and recycles red blood cells, was preprocessed by acid digestion. After a spleen sample in a homogeneous state was prepared in a 50 mL tube to a final volume of 20 mL, the level of inorganic Fe elements in the spleen was detected through ICP-MS. Furthermore, the intensity and concentration of Fe in the spleen of control mice and MNP-treated mice were identified. The intravenously injected dextran-MNPs showed the highest accumulation rate in the spleen on day 15. Therefore, the optimal period for suppressing the overactivation of T cells in the spleen is 15 days. 생체 적합성 재료로 사용되고 있는 자성나노입자는 생명체를 구성하는 14가지 미량 원소들 중 하나로서 약물전달, 발열요법 등 다양한 바이오 응용분야에 활용되고 있다. 마취가 된 마우스 꼬리 정맥에 1마리는 PBS만을 주입하고 나머지 4마리는 2 μg/μL 농도의 자성나노입자를 각각 주입하고 복부를 개복하여 총 7가지의 장기를 해부함과 동시에 중량 측정을 진행한다. 채취한 장기 중 T세포와 적혈구를 생성하는 비장만 분리하여 시료의 전 처리 과정을 수행한다. 50 mL 튜브에 균질액 상태인 비장 샘플을 최종 부피 20 mL로 만든 후 ICP-MS를 통해 시료를 구성하고 있는 성분 중 비장 내 Fe 무기원소의 수치를 검출한다. 대조군 마우스부터 20일차 마우스까지의 비장 내 Fe 강도와 비장 내 Fe 농도를 분석한다. 정맥으로 주입된 덱스트란-자성나노입자는 15일차에 비장에서 가장 높은 축적률을 보인다. 따라서 비장 내 T세포 과잉 활성화를 억제하기 위한 최적의 기간은 15일이다.