Mapping Epitopes by Phage Display.

Monoclonal antibodies (mAbs) are valuable biological molecules, serving for many applications. Therefore, it is advantageous to know the interaction pattern between antibodies and their antigens. Regions on the antigen which are recognized by the antibodies are called epitopes, and the respective molecular counterpart of the epitope on the mAbs is called paratope.

Antibody Selection via Phage Display in Microtiter Plates.

The most common and robust in vitro technology to generate monoclonal human antibodies is phage display. This technology is a widely used and powerful key technology for recombinant antibody selection. Phage display-derived antibodies are used as research tools, in diagnostic assays, and by 2022, 14 phage display-derived therapeutic antibodies were approved.

Construction of Human Immune and Naive scFv Phage Display Libraries.

Antibody phage display is a widely used in vitro selection technology for the generation of human recombinant antibodies and has yielded thousands of useful antibodies for research, diagnostics, and therapy. In order to successfully generate antibodies using phage display, the basis is the construction of high-quality antibody gene libraries. Here, we describe detailed methods for the construction of such high-quality immune and naive scFv gene libraries of human origin.

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Despite readily available alternative technologies, many antibodies in research and diagnostics are still generated and produced with the use of laboratory animals. Here, we portrait the concept of animal-free recombinant antibodies and the generation of such an antibody against the spike-protein of SARS-Cov-2. The scientific community needs to raise awareness for more sustainable animal-free alternatives, especially as they offer a path to more reliable and reproducible data.

Generation of in situ sequencing based OncoMaps to spatially resolve gene expression profiles of diagnostic and prognostic markers in breast cancer.

Background: Gene expression analysis of breast cancer largely relies on homogenized tissue samples. Due to the high degree of cellular and molecular heterogeneity of tumor tissues, bulk tissue-based analytical approaches can only provide very limited system-level information about different signaling mechanisms and cellular interactions within the complex tissue context.

Methods: We describe an analytical approach using in situ sequencing (ISS), enabling highly multiplexed, spatially and morphologically resolved gene expression profiling.

Epitope Mapping via Phage Display from Single-Gene Libraries.

Antibodies are widely used in a large variety of research applications, for diagnostics and therapy of numerous diseases, primarily cancer and autoimmune diseases. Antibodies are binding specifically to target structures (antigens). The antigen-binding properties are not only dependent on the antibody sequence, but also on the discrete antigen region recognized by the antibody (epitope).

Long-Term Follow-Up of Fundus Autofluorescence Imaging Using Wide-Field Scanning Laser Ophthalmoscopy.

Aim: To evaluate the variation of peripheral alterations in different retinal diseases over a period of >3 years by using wide-field fundus autofluorescence (FAF) scanning laser ophthalmoscopy (SLO).

Methods: A total of 26 eyes from 13 patients (median age 66 years, range 19-80) with age-related macular degeneration and other retinal degenerations were examined. In 2009, the Optos P200CAF prototype and from 2012 onwards, the Optos 200Tx (Optos plc, Scotland) were used for wide-field FAF SLO (excitation 532 nm).

Patient preference of ranibizumab treatment regimen for neovascular age-related macular degeneration - monthly injections versus pro re nata.

Purpose: To identify preference of treatment regimen in patients with anti-VEGF therapy for neovascular age-related macular degeneration (AMD) in real life.

Methods: A cross-sectional study was conducted in 200 patients receiving ranibizumab therapy on a pro re nata regimen with monthly controls. One hundred and twenty-four patients were recruited in a tertiary health care clinic, and 76 patients were recruited in a private practice.

[Breaking the line of defence].

Tarantulas have urticating hairs which may cause severe irritation of the skin, respiratory tract and eyes. Inflammation and granulomas (ophthalmia nodosa) can develop in the anterior and posterior segment of the eye presenting as keratoconjunctivitis or uveitis. We present a case of a female patient with an intracorneal tarantula hair and subsequent keratouveitis after contact with a pet tarantula.

[Isolated bilateral Sternocostoclavicular Hyperostosis (SCCH) with a pathological clavicular fracture--a 10-year follow-up].

The presented case demonstrates the clinical and radiological course of a woman suffering from bilateral sternocostoclavicular hyperostosis (SCCH) from early changes to a spontaneous clavicula fracture after 10 years. SCCH is characterised by a chronic recurrent painful swelling of the sternoclavicular region due to an aseptic inflammation and hyperostosis of the clavicula, sternum, upper ribs and the adjacent soft tissues. Frequently the picture is accompanied by cutaneous and other skeletal symptoms.

[Bacterial osteitis. Special considerations in immunocompromised patients].

With increasing life expectancy and better medical competence, the number of older patients with multimorbidity is growing. Patients with a deficient immune system need more attention during diagnosis and treatment of osteomyelitis.

[Arm motion analysis: a new method and its clinical application].

Aim: An investigation into the objective criteria of shoulder mobility possesses special meaning for diagnostic documentation, the evaluation of therapy and the effects of rehabilitation in the treatment of diseases of the shoulder. In order to ascertain the criteria which characterise shoulder mobility, it is necessary to have a tool available that is objective, comparable and allows the complexity, variability and range of motion to be recorded. Motion analysis represents one such standard procedure used to measure joint movement.

[Portable NMR-MOUSE: a new method and its evaluation of the Achilles tendon].

Aim: The increased demands placed on the Achilles tendon in recreational and professional sports have led to a rise in pathological changes in this anatomic region. Magnetic resonance evaluations of the tendon have proven to be a valid but expensive and stationary diagnostic tool. In this study, a new mobile NMR sensor was to be tested in the evaluation of the Achilles tendon.

[The value of peripheral quantitative computed tomography (pQCT) in the diagnosis of osteoporosis].

Aim: To evaluate the accuracy of distal radial pQCT in discriminating between osteoporotic fracture and non fracture cases and its efficiency to predict fractures.

Methods: Densitometric data determined with an XCT 900 Stratec pQCT device at the distal radius of women who had suffered distal radius (57), spinal (87) or femoral neck fractures (21) before or after measurement were compared with densitometric data of age-matched healthy women (265) and were analyzed in regression tests. Women whose fractures occurred before measuring were selectively analyzed.

Impact of hematocrit value after coronary artery surgery on perioperative myocardial infarction rate.

Background: The optimal hematocrit (HCT) value after coronary artery bypass grafting on cardiopulmonary bypass (CPB) has not yet been established. The purpose of our retrospective study was to investigate the association between HCr at the time of entry into the ICU and perioperative Ml rate.

Methods: We reviewed the charts of 500 consecutive coronary artery surgery patients with respect to biometric data, operative procedure, aprotinin or tranexamic acid use, perioperative drainage blood loss and transfusion requirements, perioperative Ml, ICU stay and hospital mortality.

Tibio-calcaneo-naviculo-cuboidale arthrodesis: 6 patients followed for 1-8 years.

As a salvage procedure for patients with irreparable damage to the peritalar anatomy, we used a new method, the Ilizarov ring fixator, to stabilize a tibio-calcaneo-naviculo-cuboideal (TCNC) arthrodesis after resecting the talus in 6 patients with an average follow-up of 3 years. The tibia was lengthened by callus distraction, mean 6 (3.5-10) cm.

Septic arthritis of the knee in adults: treatment by arthroscopy or arthrotomy.

Fifty-one patients with septic arthritis of the knee were reviewed retrospectively. Twenty-seven patients had been treated by arthroscopic lavage and debridement, 24 patients by open arthrotomy with subtotal synovectomy. The patients were staged according to the duration of preoperative symptoms and to the intraoperative spread of the inflammatory process.

Interleukin-6: a potential inflammatory marker after total joint replacement.

In a prospective study C-reactive protein (CRP) and interleukin-6 (IL-6) measurements were taken serially in 30 patients before and after 20 total hip arthroplasties (THR) and 10 total knee arthroplasties (TKR). There were no peri- and postoperative complications. Postoperatively the IL-6 serum concentration increased rapidly and peaked 6 h postoperatively at maximum levels (399+/-140 pg/ml).

Diagnostic and therapeutic management of lumbar and thoracic spondylodiscitis--an evaluation of 59 cases.

Fifty-nine patients with spondylodiscitis (SD) of the thoracic and/or lumbar spine were followed-up clinically and radiologically [X-ray, computed tomography (CT), magnetic resonance imaging (MRI)] over a mean time of 2.2 years (1-6.5 years).

[Early diagnosis of isthmic spondylolysis with MRI].

Introduction: Early diagnosis of isthmic lumbar spondylolysis cannot always be established on plain radiographs and CT scans, only. In the case presented here, magnetic resonance imaging (MRI) showed typical bone marrow changes in T1- and T2-weighted images, even at an early stage.

Case: A 11-year old female judoka complained of deep lumbar pain with local tenderness to pressure at L3 to S1.

[Optimizing the bone cement-implant interface by hydrolysis-resistant conditioning of the metal surface].

Purpose: The hydrolytic degradation of the implant-cement interface has to be seen as the main reason for aseptic loosening of cemented total hip replacements. Therefore, a new method of conditioning the metallic surface was developed in order to achieve a hydrolytic-resistant bound stability between the implant and bone cement. Preliminary experimental data on test bodies are presented here.

[A new MR-(and CT-) compatible bone biopsy system: first clinical results].

Purpose: To describe the development and clinical evaluation of a new, hand-powered or alternatively motor-driven, MRI and CT compatible percutaneous bone biopsy system.

Materials And Methods: A new coaxial drill system (Cook Europe A/S) was designed for percutaneous, MR-guided bone biopsies and powered either by hand or an optional motor (10-250 rotations/minute using 6 bar [88.2 PSI] compressed air).

[Percutaneous CT-controlled puncture and drainage of spondylodiscitis--a minimally invasive method].

Purpose: Is there an absolute operative indication for the abscess-forming spondylodiscitis or is a curing also possible with minimal invasive procedures?

Method: In a retrospective study over a period from 1986 to 1993, 40 patients with a spondylodiscitis of the thoracic and lumbar spine were treated and then followed up over two years in our Department of Orthopedic Surgery. Using a CT-controlled method of minimal invasive punction and drainage of the abscessed forms of spondylodiscitis, it was to be settled whether these therapeutic procedures result in a normalization of the biochemical inflammatory parameters (CRP) and in a normalization of the CT--as well as MRI-findings.

Results: 7 of the 40 patients had a spondylodiscitis with a local abscess, further 7 patients had a gravidation abscess.

[MRI-controlled outcome after core decompression of the femur head in aseptic osteonecrosis and transient bone marrow edema].

Purpose: We evaluated the signal changes of avascular necrosis and transient bone marrow oedema before and after core decompression of the femoral head to deduce prognostic factors for this operative procedure.

Method: 38 to 40 cases with core decompression (35 patients, medium age 44 years) were analysed with a mean follow-up time of 26 months. Preoperatively patients were staged by the ARCO classification.

MRI-guided biopsy of bone in a hybrid system.

This is a report of our experience with percutaneous, MRI-guided biopsies in 25 patients with skeletal lesions using a 1.5-T MR hybrid system. Twenty-five consecutive patients with skeletal lesions were referred for MRI-guided biopsy.