3-Glucuronidase from human placenta was purified 18 000-fold to homogeneity in three steps: (1) batch immunoadsorption on antibody-Sepharose resin; (2) DEAE-Sephadex chromatography; and (3) reduction and alkylation followed by DEAE-Sephadex chromatography. The product behaved as a single species when tested by (a) polyacrylamide gel electrophoresis, (b) gel filtration on Sephadex G-200, (c) sedimentation equilibrium data, and (d) double immunodiffusion against either goat or rat antibody to 3glucuronidase. The molecular weight determination gave values of 310 000 ± 10 000 (gel filtration) and 286 000 ± 10 000 (sedimentation equilibrium). Amino acid analysis showed no cysteine. No radioactivity was incorporated into the protein upon reduction and alkylation with [14C]iodoacetic ^/-Glucuronidase (EC 3.2.1.31) is a lysosomal acid hydrolase which has been studied extensively in different mammalian systems. Rat liver 3-glucuronidase (de Duve et al., 1955) and mouse liver 3-glucuronidase (Paigen, 1961;Fishman et al., 1969) are found in both lysosomes and microsomes. Mouse liver 3-glucuronidase has multiple forms (Swank and Paigen, 1973), tetrameric structure, carbohydrate residues, and only trace amounts of cysteine (Tomino et al., 1975). The 3-glucuronidase from rat liver (Stahl and Touster, 1971;Owens et al., 1975) or rat preputial gland (Himeno et al., 1974;Tulsiani et al., 1975) has chemical, physical, and biochemical properties similar to the mouse enzyme. 3-Glucuronidase from bovine liver (Plapp and Cole, 1966) was found to have multiple forms which differed in carbohydrate content and isoelectric point (Plapp and Cole, 1967). Human placental enzyme has been partially purified (Contractor and Shane, 1972;Contractor and Oakey, 1977) and the human liver enzyme purified to apparent homogeneity (Musa et al., 1965). However, the properties of the latter differ from those we report here, post From the

Neuron-specific enolase (NSE) has been increasingly recognized as a marker for neuroendocrine tumors including small cell carcinoma of the lung (SCCL). To prepare monoclonal antibodies (MAbs) specific for human NSE, we first developed a simple method of purifying NSE by direct chromatofocusing of a crude extract of human brain tissue. BALB/c mice were then immunized with our preparation of NSE, and MAbs against NSE were generated utilizing a hybridoma technique. The antibodies were screened against both NSE and non-neuronal enolase (NNE) by a solid-phase radioimmunoassay (SPRIA). After cloning and subcloning of hybridomas, two groups of anti-NSE MAbs were identified by SPRIA. One group reacted specifically with NSE but not with its isoenzyme NNE, irrespective of whether antigens were glutaraldehyde fixed or unfixed. A second group reacted with both NSE and NNE when the latter were glutaraldehyde fixed, but surprisingly with neither antigen in the absence of fixation. Group I antibodies were further characterized by immunoblotting, and by immunocytochemistry of normal brain and liver sections and sections of SCCL. The results further supported the specificity of group I antibodies for NSE. These MAbs have potential utility in the diagnosis and management of neuroendocrine tumors, and in further understanding the biology of NSE.

No abstract

The plasma membrane antigens of an oat-cell carcinoma of the lung were studied to determine if any antigens absent from normal adult tissue could be identified. Rabbit and monkey antisera were prepared to a highly purified plasma membrane fraction of an oat-cell carcinoma of the lung. The specificities of the antisera were studied by the indirect immunofluorescence method on frozen section substrate. The rabbit antiserum, after absorption with normal lung, liver, colon and peripheral nerve homogenates and extracts, failed to react with any detectable normal adult tissue. The absorbed anti-serum did react with 7 of 7 oat-cell carcinomas of the lung, but failed to react with any of 7 adeno-carcinomas of the lung, 6 epidermoid carcinomas of the lung, 7 colon carcinomas, 8 breast carcinomas, 4 kidney carcinomas, and 1 pancreatic carcinoma. The unabsorbed monkey antiserum failed to react with any detectable normal adult tissue, and had a tumor reactivity pattern nearly identical to that of the absorbed rabbit antiserum. Thus similar results were obtained with antisera from two different species. It is concluded that oat-cell carcinomas of the lung express a plasma membrane antigen(s) undetectable in normal adult tissue and highly associated with this tumor type.

Histocompatibility antigen typing was carried out in 50 Caucasian patients with primary open-angle glaucoma (POAG) and 50 Caucasian ocular-normotensive subjects. HLA-A 3 was present in 46%, B7 in 52%, B 12 in 50%, and either B 7 or B 12 in 88% of patients with POAG. These prevalences in POAG patients were significantly greater than in ocular-normotensive subjects (p < 0.01, p < 0.0005, p < 0.001, and p < 0.0005, respectively). The prevalences of A 3-B 7, A 3-B 12 and either combination were also significantly greater in POAG patients than in the ocular normotensives (p < 0.005, p < 0.005, and p < 0.0005, respectively). HLA-BW 35 was noted to be in deficit in Caucasian POAG patients (8%) as compared to Caucasian ocular normotensives (32%; p < 0.01).